Pancreatic ductal adenocarcinoma (PDAC) is distinguished by rapid dissemination. Thus, genetic and/or epigenetic deregulation of metastasis suppressor genes (MSG) is a likely event during early pancreatic carcinogenesis and a potential diagnostic marker for the disease. We investigated 9 known MSGs for their role in the dissemination of PDAC and examined their promoters for methylation and its use in PDAC detection.
A new assay for measuring chromosome instability (CIN) and identification of...Enrique Moreno Gonzalez
This document describes a new assay for measuring chromosome instability (CIN) using a human artificial chromosome (HAC) carrying a fluorescent marker. The assay allows quantification of HAC loss after drug treatment via flow cytometry. Several known anti-mitotic drugs were tested and found to increase HAC loss rates to varying degrees, with microtubule-stabilizing drugs taxol and peloruside A showing the highest increases in CIN. This new assay provides a simple and efficient way to screen drugs and identify those that elevate CIN, with the goal of developing new cancer therapies targeting chromosomally unstable tumors.
Variant G6PD levels promote tumor cell proliferation or apoptosis via the STA...Enrique Moreno Gonzalez
Glucose-6-phosphate dehydrogenase (G6PD), elevated in tumor cells, catalyzes the first reaction in the pentose-phosphate pathway. The regulation mechanism of G6PD and pathological change in human melanoma growth remains unknown.
Frizzled-8 receptor is activated by the Wnt-2 ligand in non-small cell lung c...Enrique Moreno Gonzalez
This document summarizes a research study that investigated the activation of Wnt-2 signaling through the Frizzled-8 receptor in non-small cell lung cancer (NSCLC). The study found a correlation between increased expression of Wnt-2 and Frizzled-8 in lung cancer tissue samples. A novel dominant-negative Wnt-2 construct (dnhWnt-2) inhibited Wnt-2 signaling activation and reduced colony formation of NSCLC cells in vitro and tumor growth in a mouse xenograft model. The dnhWnt-2 construct may provide a new therapeutic approach for targeting the Wnt pathway in lung cancer.
Overexpression of peptide deformylase in breast, colon, and lung cancersEnrique Moreno Gonzalez
Human mitochondrial peptide deformylase (PDF) has been proposed as a novel cancer therapeutic target. However, very little is known about its expression and regulation in human tissues. The purpose of this study was to characterize the expression pattern of PDF in cancerous tissues and to identify mechanisms that regulate its expression.
This document provides guidance on the use of Bristol-Myers Squibb (BMS) content for scientific exchange with healthcare professionals (HCPs) regarding nivolumab and metastatic renal cell carcinoma (mRCC). Key points include:
- BMS material may be used reactively for scientific exchange with HCPs and provided to HCPs requesting medical information.
- If appropriate based on country approval status, inform HCPs if information provided is off-label.
- The material was approved for use as described by BMS medical and obtained necessary permissions.
- External use requires appropriate medical, regulatory and legal review per local rules.
The document summarizes a study investigating the expression and role of apoptosis-related molecules like TRAIL, FasL, and their receptors in pancreatic adenocarcinoma cell lines. The key findings were:
1) The pancreatic cancer cell lines expressed high levels of apoptosis-inducing ligands and receptors but showed variable susceptibility to TRAIL-induced cell death.
2) Treatment with chemotherapy drugs did not increase their susceptibility to apoptosis, likely due to their differential expression of decoy receptors and inhibitor molecules.
3) This suggests pancreatic cancers develop resistance to immune-mediated apoptosis, allowing immune evasion and tumor progression.
The document discusses HER2 splice variants and their relevance in CREB1-mediated responses in breast cancer cell lines. It outlines experiments to test the effects of different HER2 variants and drug combinations on cell proliferation, invasion, CREB1 mRNA and protein levels. The methodology creates HER2 variant cell lines and tests drug combinations of Lapatinib, Trastuzumab and Pertuzumab. It is expected that the Δ16HER2+ variant will respond best to Lapatinib and Pertuzumab, while the CTF-687+ variant will respond to Lapatinib alone. The unmodified lines are expected to respond best to Lapatinib and Pertuzumab together.
1) Prodigiosin, a bacterial metabolite, induces apoptosis in human breast cancer cells. Gene expression profiling found that prodigiosin strongly increased expression of the NAG-1 gene.
2) Experiments showed that prodigiosin triggers accumulation of the tumor suppressor protein p53, but induction of NAG-1 was independent of p53.
3) Prodigiosin causes inhibition of AKT and activation of glycogen synthase kinase-3B (GSK-3B). Induction of NAG-1 and apoptosis correlated with GSK-3B activation. Inhibiting GSK-3B reduced apoptosis, suggesting GSK-3B plays a key role in the proap
A new assay for measuring chromosome instability (CIN) and identification of...Enrique Moreno Gonzalez
This document describes a new assay for measuring chromosome instability (CIN) using a human artificial chromosome (HAC) carrying a fluorescent marker. The assay allows quantification of HAC loss after drug treatment via flow cytometry. Several known anti-mitotic drugs were tested and found to increase HAC loss rates to varying degrees, with microtubule-stabilizing drugs taxol and peloruside A showing the highest increases in CIN. This new assay provides a simple and efficient way to screen drugs and identify those that elevate CIN, with the goal of developing new cancer therapies targeting chromosomally unstable tumors.
Variant G6PD levels promote tumor cell proliferation or apoptosis via the STA...Enrique Moreno Gonzalez
Glucose-6-phosphate dehydrogenase (G6PD), elevated in tumor cells, catalyzes the first reaction in the pentose-phosphate pathway. The regulation mechanism of G6PD and pathological change in human melanoma growth remains unknown.
Frizzled-8 receptor is activated by the Wnt-2 ligand in non-small cell lung c...Enrique Moreno Gonzalez
This document summarizes a research study that investigated the activation of Wnt-2 signaling through the Frizzled-8 receptor in non-small cell lung cancer (NSCLC). The study found a correlation between increased expression of Wnt-2 and Frizzled-8 in lung cancer tissue samples. A novel dominant-negative Wnt-2 construct (dnhWnt-2) inhibited Wnt-2 signaling activation and reduced colony formation of NSCLC cells in vitro and tumor growth in a mouse xenograft model. The dnhWnt-2 construct may provide a new therapeutic approach for targeting the Wnt pathway in lung cancer.
Overexpression of peptide deformylase in breast, colon, and lung cancersEnrique Moreno Gonzalez
Human mitochondrial peptide deformylase (PDF) has been proposed as a novel cancer therapeutic target. However, very little is known about its expression and regulation in human tissues. The purpose of this study was to characterize the expression pattern of PDF in cancerous tissues and to identify mechanisms that regulate its expression.
This document provides guidance on the use of Bristol-Myers Squibb (BMS) content for scientific exchange with healthcare professionals (HCPs) regarding nivolumab and metastatic renal cell carcinoma (mRCC). Key points include:
- BMS material may be used reactively for scientific exchange with HCPs and provided to HCPs requesting medical information.
- If appropriate based on country approval status, inform HCPs if information provided is off-label.
- The material was approved for use as described by BMS medical and obtained necessary permissions.
- External use requires appropriate medical, regulatory and legal review per local rules.
The document summarizes a study investigating the expression and role of apoptosis-related molecules like TRAIL, FasL, and their receptors in pancreatic adenocarcinoma cell lines. The key findings were:
1) The pancreatic cancer cell lines expressed high levels of apoptosis-inducing ligands and receptors but showed variable susceptibility to TRAIL-induced cell death.
2) Treatment with chemotherapy drugs did not increase their susceptibility to apoptosis, likely due to their differential expression of decoy receptors and inhibitor molecules.
3) This suggests pancreatic cancers develop resistance to immune-mediated apoptosis, allowing immune evasion and tumor progression.
The document discusses HER2 splice variants and their relevance in CREB1-mediated responses in breast cancer cell lines. It outlines experiments to test the effects of different HER2 variants and drug combinations on cell proliferation, invasion, CREB1 mRNA and protein levels. The methodology creates HER2 variant cell lines and tests drug combinations of Lapatinib, Trastuzumab and Pertuzumab. It is expected that the Δ16HER2+ variant will respond best to Lapatinib and Pertuzumab, while the CTF-687+ variant will respond to Lapatinib alone. The unmodified lines are expected to respond best to Lapatinib and Pertuzumab together.
1) Prodigiosin, a bacterial metabolite, induces apoptosis in human breast cancer cells. Gene expression profiling found that prodigiosin strongly increased expression of the NAG-1 gene.
2) Experiments showed that prodigiosin triggers accumulation of the tumor suppressor protein p53, but induction of NAG-1 was independent of p53.
3) Prodigiosin causes inhibition of AKT and activation of glycogen synthase kinase-3B (GSK-3B). Induction of NAG-1 and apoptosis correlated with GSK-3B activation. Inhibiting GSK-3B reduced apoptosis, suggesting GSK-3B plays a key role in the proap
The document outlines a PhD research proposal on using circulating tumor cells (CTCs) as biomarkers for renal cell carcinoma (RCC). The proposal includes 5 research projects: 1) A systematic literature review on CTCs in RCC. 2) A retrospective study analyzing CTC counts in RCC patient samples and their relationship to staging and recurrence. 3) A prospective study comparing CTC liquid biopsies to renal mass biopsies for diagnosis. 4) A randomized controlled trial comparing surgical techniques for radical nephrectomy. 5) A randomized controlled trial comparing surgical techniques for partial nephrectomy. Detailed methods, timelines and practical considerations are provided for each project.
ADAR2 editing activity in newly diagnosed versus relapsed pediatric high-grad...Enrique Moreno Gonzalez
High-grade (WHO grade III and IV) astrocytomas are aggressive malignant brain tumors affecting humans with a high risk of recurrence in both children and adults. To date, limited information is available on the genetic and molecular alterations important in the onset and progression of pediatric high-grade astrocytomas and, even less, on the prognostic factors that influence long-term outcome in children with recurrence. A-to-I RNA editing is an essential post-transcriptional mechanism that can alter the nucleotide sequence of several RNAs and is
mediated by the ADAR enzymes. ADAR2 editing activity is particularly important in mammalian brain and is impaired in both adult and pediatric high-grade astrocytomas.
Moreover, we have recently shown that the recovered ADAR2 activity in high-grade astrocytomas inhibits in vivo tumor growth. The aim of the present study is to investigate whether changes may occur in ADAR2-mediated RNA editing profiles of relapsed highgrade astrocytomas compared to their respective specimens collected at diagnosis, in four pediatric patients.
The document summarizes research on using a small molecule mimetic of the Sin3A interaction domain (SID) decoy (C16) alone or in combination with retinoic acid receptor alpha (RARα) agonists to treat triple negative breast cancer (TNBC). The key findings are:
1) C16 increases expression of RARβ2 and endogenous retinoic acid levels, activating RAR target gene expression.
2) C16 combined with the RARα agonist AM80 strongly activates RARE-driven reporter gene expression more than either treatment alone.
3) In 3D culture, C16 combined with RARα agonists induces acinar morphogenesis and inhibits tumors
This document summarizes a study investigating the role of microRNA-122 (miR-122) in regulating intrahepatic metastasis of hepatocellular carcinoma (HCC). The study found that miR-122 expression is significantly downregulated in HCC tumors with intrahepatic metastasis. Restoring miR-122 expression in metastatic HCC cell lines reduced in vitro migration, invasion, and tumor growth in vivo. Computational analysis identified multiple target genes of miR-122, including ADAM17, which was shown to be involved in metastasis. Silencing ADAM17 had similar effects as restoring miR-122, reducing in vitro and in vivo measures of metastasis. The study suggests that miR-122 acts as a tumor suppressor
OSU-03012 sensitizes breast cancers to lapatinib-induced cell killing: a role...Enrique Moreno Gonzalez
Lapatinib is characterized as an ErbB1/ErbB2 dual inhibitor and has recently been approved for the treatment of metastatic breast cancer. In this study, we examined mechanisms
associated with enhancing the activity of lapatinib via combination with other therapies.
The document analyzes the tumor transcriptomics profile of a patient with stage IV gallbladder cancer. It identifies the top 200 most significantly activated expression regulators in the patient's tumor using sub-network enrichment analysis software. Key regulators identified include histone deacetylases and DNA methyltransferases. The analysis suggests treatment with the HDAC inhibitor vorinostat and discusses how activated regulators like HDACs, PDCD1, and CTLA4 may be contributing to tumor proliferation and immune evasion. Graphical summaries show pathways enriched with active regulators in the tumor related to cancer hallmarks like histone modification and immune response evasion.
Poster presentation at the 2016 WORLD GASTROINTESTINAL SYMPOSIUM on tepotinib a selective inhibitor of c-MET by S. Faivre, J.-F. Blanc, P. Merle, A. Fasolo, A. Iacobellis, V. Grando, T. Decaens, J. Trojan, E. Villa, U. Stammberger, R. Bruns, E. Raymond
1Oncology Unit, Beaujon University Hospital, Clichy, France; 2Service d’hépato-gastroentérologie et d’oncologie digestive, Groupe Hospitalier Saint André, Bordeaux, France; 3Service d'Hépato-Gastro-Entérologie, Hôpital de la Croix Rousse, Lyon, France; 4Dipartimento di Oncologia Medica, Ospedale San Raffaele IRCSS, Milan, Italy; 5Servizio di Endoscopia Digestiva, Ospedale Casa Sollievo della Sofferenza IRCCS, San Giovanni Rotondo, Italy; 6Service Hépatologie, Hôpital Jean-Verdier, Bondy, France; 7Service d'hepato-gastro-enterologie, CHU de Grenoble - Hôpital Nord, Grenoble, France; 8Gastrointestinal Oncology, Goethe University Hospital, Frankfurt, Germany; 9Policlinico di Modena, Modena, Italy; 10Merck KGaA, Darmstadt, Germany
3-Formylchromone (3-FC) is a synthetic compound that may have anti-cancer effects through suppression of the STAT3 signaling pathway in hepatocellular carcinoma (HCC) cells. The study investigated the effects of 3-FC on STAT3 activation and the expression of STAT3-regulated genes involved in cell proliferation and survival in HCC cells. The results showed that 3-FC inhibited STAT3 activation by suppressing upstream kinases like JAK1 and JAK2. It also downregulated cyclin D1, Bcl-2, Bcl-xL and other genes regulated by STAT3. Furthermore, 3-FC treatment inhibited HCC cell proliferation and induced apoptosis. Therefore, 3
Abnormal expression of Pygopus 2 correlates with a malignant phenotype in hum...Enrique Moreno Gonzalez
Pygopus 2 (Pygo2) is a Pygo family member and an important component of the Wnt signaling transcriptional complex. Despite this data, no clinical studies investigating Pygo2 expression in lung cancer have yet been reported.
Antioxidant-mediated up-regulation of OGG1 via NRF2 induction is associated ...Enrique Moreno Gonzalez
Estrogen metabolism-mediated oxidative stress is suggested to play an important role in estrogen-induced breast carcinogenesis. We have earlier demonstrated that antioxidants,
vitamin C (Vit C) and butylated hydroxyanisole (BHA) inhibit 17β-estradiol (E2)-mediated oxidative stress and oxidative DNA damage, and breast carcinogenesis in female August
Copenhagen Irish (ACI) rats. The objective of the present study was to characterize the mechanism by which above antioxidants prevent DNA damage during breast carcinogenesis.
Calcarea carbonica induces apoptosis in cancer cells in p53-dependent manner ...home
These observations delineate the significance of immuno-modulatory circuit during calcarea carbonicamediated
tumor apoptosis. The molecular mechanism identified may serve as a platform for involving calcarea
carbonica into immunotherapeutic strategies for effective tumor regression
Angiogenesis in the compound literature Cell Cycle andcross-screening studies.dish33pest
The document discusses MAGE-A4, a protein involved in angiogenesis. It finds that lower doses of Adriamycin increase proteasome activity and promote the proteasome-mediated processing of MAGE-A4. Specifically, it shows that Adriamycin, but not other anticancer drugs, increases the processed form of MAGE-A4 under the conditions tested. The study analyzed the mechanism of MAGE-A4 processing and found that Adriamycin selectively increases proteasome-mediated cleavage of MAGE-A4 compared to other anticancer agents.
1) The study found that the tumor protein p53 regulates the expression of the immune checkpoint protein PDL1 (programmed death ligand 1) via the microRNA miR-34.
2) Delivery of miR-34a in a mouse model of lung cancer using a liposomal formulation (MRX34) reduced PDL1 expression in tumors and increased tumor-infiltrating immune cells.
3) Combining MRX34 with radiation therapy further increased immune cell numbers in tumors and showed potential as a novel cancer therapeutic approach.
This paper reports phase II clinical data on galunisertib, a novel small molecule TGF-beta receptor inhibitor in patients with advanced non-operable hepatocellular carcinoma. This poster was presented at ASCO 2016 in Chicago. Results show activity and mild toxicity.
This study identified a recurrent amplification of the VEGFA gene locus in a subset of mouse and human hepatocellular carcinomas (HCC). Tumors with this amplification (Amppos) displayed distinct characteristics including higher proliferation, vessel density, and macrophage content compared to tumors without the amplification (Ampneg). Additionally, Amppos tumors had increased expression of VEGFA and other genes in the amplified region. Treatment with sorafenib, which targets VEGFR, was found to be particularly effective in inhibiting the growth of Amppos HCCs both in mouse models and human patients. This suggests VEGFA amplification could serve as a biomarker to identify HCC patients most likely to benefit from sorafenib or other
This study developed a new mouse model that lacks all isoforms of the Trim24 gene. Mice with complete loss of Trim24 (Trim24-/-) spontaneously developed hepatic lipid accumulation, inflammation, fibrosis, and hepatocellular carcinoma without a high-fat diet or other induction methods. Trim24-/- mice had decreased expression of genes involved in oxidation, lipid metabolism, and increased expression of genes related to endoplasmic reticulum stress and cell cycle pathways. By 6 months of age, Trim24-/- mice exhibited macroscopic white liver lesions composed of steatosis and fibrosis that progressed to hepatocellular carcinoma, recapitulating features of non-alcoholic fatty liver disease in humans
1) The document discusses a recent study that found focal amplifications in the VEGFA gene occur in 7-10% of hepatocellular carcinoma (HCC) cases. 2) The study showed that in HCC tumors with VEGFA amplification, VEGF-A stimulates the secretion of hepatocyte growth factor (HGF) by stromal cells, promoting tumor progression. 3) HCC patients with VEGFA amplification seemed to have a better response to the drug sorafenib, suggesting VEGFA amplification could predict sorafenib effectiveness.
This document discusses evaluating cancer testis antigens (CTAs) as targets for immunotherapy of triple negative breast cancer (TNBC). It finds that the gene expression of DKKL1, LDHC, MAGE-A3, PIWIL2, PLAC1, PRAME, PRSS50 and TSGA10 are moderately or highly expressed in human TNBC cell lines and datasets, making them potential immunotherapeutic targets. Real-time qRT-PCR and western blotting confirmed expression of some CTAs at the gene and protein level in the cell lines.
Purpose: An inherited mutation in KRAS (LCS6-variant or rs61764370) results in altered control of the KRAS oncogene. We studied this biomarker’s correlation to anti-EGFR monoclonal antibody (mAb) therapy
response in patients with metastatic colorectal cancer.
Experimental Design: LCS6-variant and KRAS/BRAF mutational status was determined in 512 patients
with metastatic colorectal cancer treated with salvage anti-EGFR mAb therapy, and findings correlated with
outcome. Reporters were tested in colon cancer cell lines to evaluate the differential response of the LCS6-
variant allele to therapy exposure.
Results: In this study, 21.2% (109 of 512) of patients with metastatic colorectal cancer had the LCS6-
variant (TG/GG), which was found twice as frequently in the BRAF-mutated versus the wild-type (WT) group
(P = 0.03). LCS6-variant patients had significantly longer progression- free survival (PFS) with anti-EGFR
mAb monotherapy treatment in the whole cohort (16.85 vs. 7.85 weeks; P = 0.019) and in the double WT
(KRAS and BRAF) patient population (18 vs. 10.4 weeks; P = 0.039). Combination therapy (mAbs plus
chemotherapy) led to improved PFS and overall survival (OS) for nonvariant patients, and brought their
outcome to levels comparable with LCS6-variant patients receiving anti-EGFR mAb monotherapy. Combination
therapy did not lead to improved PFS or OS for LCS6-variant patients. Cell line studies confirmed a
unique response of the LCS6-variant allele to both anti-EGFR mAb monotherapy and chemotherapy.
Conclusions: LCS6-variant patients with metastatic colorectal cancer have an excellent response to anti-EGFR
mAb monotherapy, without any benefit from the addition of chemotherapy. These findings further confirm
the importance of thismutation as a biomarker of anti-EGFR mAb response in patients with metastatic colorectal cancer, and warrant further prospective confirmation.
Connexin-43 can delay early recurrence and metastasis in patients with hepati...Enrique Moreno Gonzalez
We studied the relationships among Cx43, CD105, and VEGF in specimens of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) with different serum AFP levels with respect to recurrence and metastasis.
Low expression of N-myc downstream-regulated gene 2 in oesophageal squamous c...Enrique Moreno Gonzalez
It is currently unclear whether a correlation exists between N-myc downstream-regulated gene 2 (NDRG2) expression and oesophageal squamous cell carcinoma (ESCC). The aim of this study was to examine the underlying clinical significance of NDRG2 expression in ESCC patients and to investigate the effects of NDRG2 up-regulation on ESCC cell growth in vitro and in vivo.
The document outlines a PhD research proposal on using circulating tumor cells (CTCs) as biomarkers for renal cell carcinoma (RCC). The proposal includes 5 research projects: 1) A systematic literature review on CTCs in RCC. 2) A retrospective study analyzing CTC counts in RCC patient samples and their relationship to staging and recurrence. 3) A prospective study comparing CTC liquid biopsies to renal mass biopsies for diagnosis. 4) A randomized controlled trial comparing surgical techniques for radical nephrectomy. 5) A randomized controlled trial comparing surgical techniques for partial nephrectomy. Detailed methods, timelines and practical considerations are provided for each project.
ADAR2 editing activity in newly diagnosed versus relapsed pediatric high-grad...Enrique Moreno Gonzalez
High-grade (WHO grade III and IV) astrocytomas are aggressive malignant brain tumors affecting humans with a high risk of recurrence in both children and adults. To date, limited information is available on the genetic and molecular alterations important in the onset and progression of pediatric high-grade astrocytomas and, even less, on the prognostic factors that influence long-term outcome in children with recurrence. A-to-I RNA editing is an essential post-transcriptional mechanism that can alter the nucleotide sequence of several RNAs and is
mediated by the ADAR enzymes. ADAR2 editing activity is particularly important in mammalian brain and is impaired in both adult and pediatric high-grade astrocytomas.
Moreover, we have recently shown that the recovered ADAR2 activity in high-grade astrocytomas inhibits in vivo tumor growth. The aim of the present study is to investigate whether changes may occur in ADAR2-mediated RNA editing profiles of relapsed highgrade astrocytomas compared to their respective specimens collected at diagnosis, in four pediatric patients.
The document summarizes research on using a small molecule mimetic of the Sin3A interaction domain (SID) decoy (C16) alone or in combination with retinoic acid receptor alpha (RARα) agonists to treat triple negative breast cancer (TNBC). The key findings are:
1) C16 increases expression of RARβ2 and endogenous retinoic acid levels, activating RAR target gene expression.
2) C16 combined with the RARα agonist AM80 strongly activates RARE-driven reporter gene expression more than either treatment alone.
3) In 3D culture, C16 combined with RARα agonists induces acinar morphogenesis and inhibits tumors
This document summarizes a study investigating the role of microRNA-122 (miR-122) in regulating intrahepatic metastasis of hepatocellular carcinoma (HCC). The study found that miR-122 expression is significantly downregulated in HCC tumors with intrahepatic metastasis. Restoring miR-122 expression in metastatic HCC cell lines reduced in vitro migration, invasion, and tumor growth in vivo. Computational analysis identified multiple target genes of miR-122, including ADAM17, which was shown to be involved in metastasis. Silencing ADAM17 had similar effects as restoring miR-122, reducing in vitro and in vivo measures of metastasis. The study suggests that miR-122 acts as a tumor suppressor
OSU-03012 sensitizes breast cancers to lapatinib-induced cell killing: a role...Enrique Moreno Gonzalez
Lapatinib is characterized as an ErbB1/ErbB2 dual inhibitor and has recently been approved for the treatment of metastatic breast cancer. In this study, we examined mechanisms
associated with enhancing the activity of lapatinib via combination with other therapies.
The document analyzes the tumor transcriptomics profile of a patient with stage IV gallbladder cancer. It identifies the top 200 most significantly activated expression regulators in the patient's tumor using sub-network enrichment analysis software. Key regulators identified include histone deacetylases and DNA methyltransferases. The analysis suggests treatment with the HDAC inhibitor vorinostat and discusses how activated regulators like HDACs, PDCD1, and CTLA4 may be contributing to tumor proliferation and immune evasion. Graphical summaries show pathways enriched with active regulators in the tumor related to cancer hallmarks like histone modification and immune response evasion.
Poster presentation at the 2016 WORLD GASTROINTESTINAL SYMPOSIUM on tepotinib a selective inhibitor of c-MET by S. Faivre, J.-F. Blanc, P. Merle, A. Fasolo, A. Iacobellis, V. Grando, T. Decaens, J. Trojan, E. Villa, U. Stammberger, R. Bruns, E. Raymond
1Oncology Unit, Beaujon University Hospital, Clichy, France; 2Service d’hépato-gastroentérologie et d’oncologie digestive, Groupe Hospitalier Saint André, Bordeaux, France; 3Service d'Hépato-Gastro-Entérologie, Hôpital de la Croix Rousse, Lyon, France; 4Dipartimento di Oncologia Medica, Ospedale San Raffaele IRCSS, Milan, Italy; 5Servizio di Endoscopia Digestiva, Ospedale Casa Sollievo della Sofferenza IRCCS, San Giovanni Rotondo, Italy; 6Service Hépatologie, Hôpital Jean-Verdier, Bondy, France; 7Service d'hepato-gastro-enterologie, CHU de Grenoble - Hôpital Nord, Grenoble, France; 8Gastrointestinal Oncology, Goethe University Hospital, Frankfurt, Germany; 9Policlinico di Modena, Modena, Italy; 10Merck KGaA, Darmstadt, Germany
3-Formylchromone (3-FC) is a synthetic compound that may have anti-cancer effects through suppression of the STAT3 signaling pathway in hepatocellular carcinoma (HCC) cells. The study investigated the effects of 3-FC on STAT3 activation and the expression of STAT3-regulated genes involved in cell proliferation and survival in HCC cells. The results showed that 3-FC inhibited STAT3 activation by suppressing upstream kinases like JAK1 and JAK2. It also downregulated cyclin D1, Bcl-2, Bcl-xL and other genes regulated by STAT3. Furthermore, 3-FC treatment inhibited HCC cell proliferation and induced apoptosis. Therefore, 3
Abnormal expression of Pygopus 2 correlates with a malignant phenotype in hum...Enrique Moreno Gonzalez
Pygopus 2 (Pygo2) is a Pygo family member and an important component of the Wnt signaling transcriptional complex. Despite this data, no clinical studies investigating Pygo2 expression in lung cancer have yet been reported.
Antioxidant-mediated up-regulation of OGG1 via NRF2 induction is associated ...Enrique Moreno Gonzalez
Estrogen metabolism-mediated oxidative stress is suggested to play an important role in estrogen-induced breast carcinogenesis. We have earlier demonstrated that antioxidants,
vitamin C (Vit C) and butylated hydroxyanisole (BHA) inhibit 17β-estradiol (E2)-mediated oxidative stress and oxidative DNA damage, and breast carcinogenesis in female August
Copenhagen Irish (ACI) rats. The objective of the present study was to characterize the mechanism by which above antioxidants prevent DNA damage during breast carcinogenesis.
Calcarea carbonica induces apoptosis in cancer cells in p53-dependent manner ...home
These observations delineate the significance of immuno-modulatory circuit during calcarea carbonicamediated
tumor apoptosis. The molecular mechanism identified may serve as a platform for involving calcarea
carbonica into immunotherapeutic strategies for effective tumor regression
Angiogenesis in the compound literature Cell Cycle andcross-screening studies.dish33pest
The document discusses MAGE-A4, a protein involved in angiogenesis. It finds that lower doses of Adriamycin increase proteasome activity and promote the proteasome-mediated processing of MAGE-A4. Specifically, it shows that Adriamycin, but not other anticancer drugs, increases the processed form of MAGE-A4 under the conditions tested. The study analyzed the mechanism of MAGE-A4 processing and found that Adriamycin selectively increases proteasome-mediated cleavage of MAGE-A4 compared to other anticancer agents.
1) The study found that the tumor protein p53 regulates the expression of the immune checkpoint protein PDL1 (programmed death ligand 1) via the microRNA miR-34.
2) Delivery of miR-34a in a mouse model of lung cancer using a liposomal formulation (MRX34) reduced PDL1 expression in tumors and increased tumor-infiltrating immune cells.
3) Combining MRX34 with radiation therapy further increased immune cell numbers in tumors and showed potential as a novel cancer therapeutic approach.
This paper reports phase II clinical data on galunisertib, a novel small molecule TGF-beta receptor inhibitor in patients with advanced non-operable hepatocellular carcinoma. This poster was presented at ASCO 2016 in Chicago. Results show activity and mild toxicity.
This study identified a recurrent amplification of the VEGFA gene locus in a subset of mouse and human hepatocellular carcinomas (HCC). Tumors with this amplification (Amppos) displayed distinct characteristics including higher proliferation, vessel density, and macrophage content compared to tumors without the amplification (Ampneg). Additionally, Amppos tumors had increased expression of VEGFA and other genes in the amplified region. Treatment with sorafenib, which targets VEGFR, was found to be particularly effective in inhibiting the growth of Amppos HCCs both in mouse models and human patients. This suggests VEGFA amplification could serve as a biomarker to identify HCC patients most likely to benefit from sorafenib or other
This study developed a new mouse model that lacks all isoforms of the Trim24 gene. Mice with complete loss of Trim24 (Trim24-/-) spontaneously developed hepatic lipid accumulation, inflammation, fibrosis, and hepatocellular carcinoma without a high-fat diet or other induction methods. Trim24-/- mice had decreased expression of genes involved in oxidation, lipid metabolism, and increased expression of genes related to endoplasmic reticulum stress and cell cycle pathways. By 6 months of age, Trim24-/- mice exhibited macroscopic white liver lesions composed of steatosis and fibrosis that progressed to hepatocellular carcinoma, recapitulating features of non-alcoholic fatty liver disease in humans
1) The document discusses a recent study that found focal amplifications in the VEGFA gene occur in 7-10% of hepatocellular carcinoma (HCC) cases. 2) The study showed that in HCC tumors with VEGFA amplification, VEGF-A stimulates the secretion of hepatocyte growth factor (HGF) by stromal cells, promoting tumor progression. 3) HCC patients with VEGFA amplification seemed to have a better response to the drug sorafenib, suggesting VEGFA amplification could predict sorafenib effectiveness.
This document discusses evaluating cancer testis antigens (CTAs) as targets for immunotherapy of triple negative breast cancer (TNBC). It finds that the gene expression of DKKL1, LDHC, MAGE-A3, PIWIL2, PLAC1, PRAME, PRSS50 and TSGA10 are moderately or highly expressed in human TNBC cell lines and datasets, making them potential immunotherapeutic targets. Real-time qRT-PCR and western blotting confirmed expression of some CTAs at the gene and protein level in the cell lines.
Purpose: An inherited mutation in KRAS (LCS6-variant or rs61764370) results in altered control of the KRAS oncogene. We studied this biomarker’s correlation to anti-EGFR monoclonal antibody (mAb) therapy
response in patients with metastatic colorectal cancer.
Experimental Design: LCS6-variant and KRAS/BRAF mutational status was determined in 512 patients
with metastatic colorectal cancer treated with salvage anti-EGFR mAb therapy, and findings correlated with
outcome. Reporters were tested in colon cancer cell lines to evaluate the differential response of the LCS6-
variant allele to therapy exposure.
Results: In this study, 21.2% (109 of 512) of patients with metastatic colorectal cancer had the LCS6-
variant (TG/GG), which was found twice as frequently in the BRAF-mutated versus the wild-type (WT) group
(P = 0.03). LCS6-variant patients had significantly longer progression- free survival (PFS) with anti-EGFR
mAb monotherapy treatment in the whole cohort (16.85 vs. 7.85 weeks; P = 0.019) and in the double WT
(KRAS and BRAF) patient population (18 vs. 10.4 weeks; P = 0.039). Combination therapy (mAbs plus
chemotherapy) led to improved PFS and overall survival (OS) for nonvariant patients, and brought their
outcome to levels comparable with LCS6-variant patients receiving anti-EGFR mAb monotherapy. Combination
therapy did not lead to improved PFS or OS for LCS6-variant patients. Cell line studies confirmed a
unique response of the LCS6-variant allele to both anti-EGFR mAb monotherapy and chemotherapy.
Conclusions: LCS6-variant patients with metastatic colorectal cancer have an excellent response to anti-EGFR
mAb monotherapy, without any benefit from the addition of chemotherapy. These findings further confirm
the importance of thismutation as a biomarker of anti-EGFR mAb response in patients with metastatic colorectal cancer, and warrant further prospective confirmation.
Connexin-43 can delay early recurrence and metastasis in patients with hepati...Enrique Moreno Gonzalez
We studied the relationships among Cx43, CD105, and VEGF in specimens of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) with different serum AFP levels with respect to recurrence and metastasis.
Low expression of N-myc downstream-regulated gene 2 in oesophageal squamous c...Enrique Moreno Gonzalez
It is currently unclear whether a correlation exists between N-myc downstream-regulated gene 2 (NDRG2) expression and oesophageal squamous cell carcinoma (ESCC). The aim of this study was to examine the underlying clinical significance of NDRG2 expression in ESCC patients and to investigate the effects of NDRG2 up-regulation on ESCC cell growth in vitro and in vivo.
Secular trends of salted fish consumption and nasopharyngeal carcinoma: a mul...Enrique Moreno Gonzalez
This document summarizes a study that examined the relationship between salted fish consumption and nasopharyngeal carcinoma (NPC) incidence and mortality rates in 8 regions over several decades. The study found decreasing NPC rates in Hong Kong correlated with decreasing salted fish consumption. However, these correlations were no longer significant after adjusting for changes in tobacco use and vegetable consumption. Additionally, no consistent relationships were observed between NPC rates and salted fish intake in the 7 other regions studied. The results do not strongly support the role of salted fish in explaining global NPC rate trends.
Differences in microRNA expression during tumor development in the transition...Enrique Moreno Gonzalez
The prostate is divided into three glandular zones, the peripheral zone (PZ), the transition zone (TZ), and the central zone. Most prostate tumors arise in the peripheral zone (70-75%) and in the transition zone (20-25%) while only 10% arise in the central zone. The aim of this study was to investigate if differences in miRNA expression could be a possible explanation for the difference in propensity of tumors in the zones of the prostate.
Anti-lymphangiogenic properties of mTOR inhibitors in head and neck squamous ...Enrique Moreno Gonzalez
Tumor dissemination to cervical lymph nodes via lymphatics represents the first step in the metastasis of head and neck squamous cell carcinoma (HNSCC) and is the most significant predictor of tumor recurrence decreasing survival by 50%. The lymphatic suppressing properties of mTOR inhibitors are not yet well understood.
The life in sight application study (LISA): design of a randomized controlled...Enrique Moreno Gonzalez
It is widely recognized that spiritual care plays an important role in physical and psychosocial well-being of cancer patients, but there is little evidence based research on the effects of spiritual care. We will conduct a randomized controlled trial on spiritual care using a brief structured interview scheme supported by an e-application. The aim is to examine whether an assisted reflection on life events and ultimate life goals can improve quality of life of cancer patients.
Clinical and experimental studies regarding the expression and diagnostic val...Enrique Moreno Gonzalez
Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a multifunctional Ig-like cell adhesion molecule that has a wide range of biological functions. According to previous reports, serum CEACAM1 is dysregulated in different malignant tumours and associated with tumour progression. However, the serum CEACAM1 expression in nonsmall-cell lung carcinomas (NSCLC) is unclear. The different expression ratio of CEACAM1-S and CEACAM1-L isoform has seldom been investigated in NSCLC. This research is intended to study the serum CEACAM1 and the ratio of CEACAM1-S/L isoforms in NSCLC.
paraphrase the review in your own wordsThe tumor suppressor PTEN .pdfarihantgiftgallery
paraphrase the review in your own words?
The tumor suppressor PTEN (phosphatase and tensin homolog deleted from chromosome 10), is
a lipid phosphatase that converts phosphatidylinositol-3, 4, 5- triphosphate (PIP3) into
phosphatidylinositol (4, 5)- diphosphate (PIP2). PTEN is well-known as the most highly mutated
tumor suppressor gene in the p53-post era [66, 67]. Recently, three papers by Nadav Bar and
Rivka Dikstein, Linhua Liu and colleagues, and Laura Poliseno and colleagues, have
demonstrated that PTEN was a bona fide target of miR-22 in a small cohort of cancer cell lines
that are driven from breast cancer, cervical cancer, prostate cancer, and bronchial epithelial
cancer. These studies contradict a uniform role of miR-22, indicating that under certain
circumstances, miR-22 may function as an oncogene because of its antagonistic effects on tumor
suppressive PTEN signaling [26, 68, 69] (Fig. 3). Using various miRNA target prediction
programs and/or RNAhybrid program for evaluating the minimum free energy hybridization,
these three groups all found that miR-22-PTEN was a high scoring miRNA-target pair. Enforced
or reduced expression of miR-22 in human HEK293T, cervical cancer HeLa and breast cancer
MCF-7 cell lines (Nadav Bar and Rivka Dikstein), anti-benzo[a]pyrene-7, 8-diol-9, 10-epoxide
(anti-BPDE)-induced transformed human bronchial epithelial cancer cell line 16HBE-T (Linhua
Liu and colleagues) and prostate cancer cell line DU145 (Laura Poliseno and colleagues),
revealed that miR-22 negatively regulated PTEN protein expression. Intriguingly, an inverse
correlation between miR-22 and PTEN mRNA expression has been presented by Poliseno et al.,
while Liu et al. found that there was no change of PTEN mRNA expression regardless of miR-
22 levels. A similarly inverse association of miR-22 and J. Xiong PTEN protein levels was
observed in 16HBE-T and its parental normal cell line16HBE (Linhua Liu and colleagues), and
in several prostate cancer cell lines, prostate cell lines and a prostate tumor tissue microarray
(Laura Poliseno and colleagues). Furthermore, the mature levels of miR-22 were significantly
increased in these tumor cells versus their normal counterparts. In line with this result, a direct
correlation between miR-22 expression and phosphorylated AKT or between the expression of
miR- 22 and that of DICER was also identified by Laura Poliseno and colleagues. These three
groups all showed that an intact binding site at the 3’UTR of PTEN mRNA was required for
miR-22 targeting. Functional analyses showed that miR-22 could induce apoptosis, inhibited
colony formation and suppressed motility of bronchial epithelial cancer cells (Linhua Liu and
colleagues), and intrinsically promote prostate cancer cell growth and tumorigenesis in tumor-
bearing nude mice (Laura Poliseno and colleagues). Subsequently, the influence of miR-22 on
the downstream signaling of PTEN was tested. Bar et al. showed that miR-22 could stimulate
AKT activity, and i.
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...JohnJulie1
Lung cancer is the second common malignancy and the most aggressive cancer worldwide with late diagnosis and poor prognosis. The search for biomarkers that promote early diagnosis and improve therapeutic strategies focuses to the understanding of the mechanisms underlying cancer development and progression. The deregulation of gene expression is one of the cancer hallmarks reflected to the stability...
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...AnonIshanvi
Lung cancer is the second common malignancy and the most aggressive cancer worldwide with late diagnosis and poor prognosis. The search for biomarkers that promote early diagnosis and improve therapeutic strategies focuses to the understanding of the mechanisms underlying cancer development and progression
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...daranisaha
Lung cancer is the second common malignancy and the most aggressive cancer worldwide with late diagnosis and poor prognosis. The search for biomarkers that promote early diagnosis and improve therapeutic strategies focuses to the understanding of the mechanisms underlying cancer development and progression. The deregulation of gene expression is one of the cancer hallmarks reflected to the stability...
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...EditorSara
Lung cancer is the second common malignancy and the most aggressive cancer worldwide with late diagnosis and poor prognosis. The search for biomarkers that promote early diagnosis and improve therapeutic strategies focuses to the understanding of the mechanisms underlying cancer development and progression. The deregulation of gene expression is one of the cancer hallmarks reflected to the stability..
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...semualkaira
Lung cancer is the second common malignancy and the most aggressive cancer worldwide with late diagnosis and poor prognosis. The search for biomarkers that promote early diagnosis and improve therapeutic strategies focuses to the understanding of the mechanisms underlying cancer development and progression. The deregulation of gene expression is one of the cancer hallmarks reflected to the stability..
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...semualkaira
Lung cancer is the second common malignancy and the most aggressive cancer worldwide with late diagnosis and poor prognosis. The search for biomarkers that promote early diagnosis and improve therapeutic strategies focuses to the understanding of the mechanisms underlying cancer development and progression. The deregulation of gene expression is one of the cancer hallmarks reflected to the stability...
Deadenylase Expression in Small Cell Lung Cancer Related To Clinical Characte...NainaAnon
This study examined the expression levels of deadenylases in small cell lung cancer (SCLC) clinical samples and their correlation to patient clinical characteristics and survival. Real-time PCR analysis found that the deadenylases PARN, CNOT6, CNOT7, and NOC were differentially expressed between malignant and normal lung tissue samples. Higher expression levels of PARN, CNOT6, and CNOT7 correlated with older patient age and decreased overall survival of 180 days or less. The results suggest these deadenylases may serve as prognostic biomarkers for SCLC patient outcomes.
Proteogenomic analysis of human colon cancer reveals new therapeutic opportun...Gul Muneer
We performed the first proteogenomic study on a prospectively collected colon cancer cohort. Comparative proteomic and phosphoproteomic analysis of paired tumor and normal adjacent tissues produced a catalog of colon cancer-associated proteins and phosphosites, including known and putative new biomarkers, drug targets, and cancer/testis antigens. Proteogenomic integration not only prioritized genomically inferred targets, such as copy-number drivers and mutation-derived neoantigens, but also yielded novel findings. Phosphoproteomics data associated Rb phosphorylation with increased proliferation and decreased apoptosis in colon cancer, which explains why this classical tumor suppressor is amplified in colon tumors and suggests a rationale for targeting Rb phosphorylation in colon cancer. Proteomics identified an association between decreased CD8 T cell infiltration and increased glycolysis in microsatellite instability-high (MSI-H) tumors, suggesting glycolysis as a potential target to overcome the resistance of MSI-H tumors to immune checkpoint blockade. Proteogenomics presents new avenues for biological discoveries and therapeutic development.
This study assessed the effects of compound A (CpdA) on bladder cancer cell growth. CpdA functions as both a glucocorticoid receptor (GR) modulator and androgen receptor (AR) antagonist. The study found that in GR/AR-positive bladder cancer cells, CpdA strongly inhibited cell proliferation, colony formation, migration and invasion. It did so predominantly by inducing GR transrepression and down-regulating nuclear factor-κB and related molecules, though it also partially acted through the AR pathway. In mouse models, CpdA more strongly suppressed tumor growth than drugs targeting only GR or AR. Thus, CpdA inhibits bladder cancer growth mainly via GR transrepression and has effects that
This study assessed the effects of compound A (CpdA) on bladder cancer cell growth. CpdA functions as both a glucocorticoid receptor (GR) modulator and androgen receptor (AR) antagonist. In GR/AR-positive bladder cancer cells, CpdA strongly inhibited cell proliferation, colony formation, migration and invasion. CpdA decreased the viability of control cells more than GR or AR silencing cells. In mouse xenograft models, CpdA more strongly suppressed tumor growth than dexamethasone or hydroxyflutamide. CpdA likely inhibits bladder cancer growth predominantly by inducing GR transrepression and partially through the AR pathway.
This document discusses a study analyzing the distribution of promoter hypermethylation of the p16 gene in male and female colorectal cancer patients in Kashmir, India. The study found different p16 promoter hypermethylation profiles in male and female patients, with occurrence being more frequent in males than females, though not statistically significant. Promoter hypermethylation of p16 was significantly associated with colorectal cancer in both sexes. The study aimed to contribute to understanding the role of epigenetic changes in colorectal cancer development and progression.
Compound A inhibits bladder cancer growth through multiple mechanisms:
1) It functions as both a glucocorticoid receptor agonist and androgen receptor antagonist, inhibiting proliferation and inducing apoptosis in bladder cancer cells expressing both receptors.
2) It promotes interactions between the glucocorticoid receptor and nuclear factor-κB, inducing glucocorticoid receptor-mediated transrepression of genes related to invasion, migration and inflammation.
3) In mouse models, Compound A more strongly suppresses tumor growth than dexamethasone or hydroxyflutamide alone, indicating its dual receptor effects are more beneficial than targeting individual receptors.
Molecular mechanisms of action and potential biomarkers of growth inhibition ...Enrique Moreno Gonzalez
Molecular targeted therapy has emerged as a promising treatment of Hepatocellular carcinoma (HCC). One potential target is the Src family Kinase (SFK). C-Src, a non-receptor tyrosine kinase is a critical link of multiple signal pathways that regulate proliferation, invasion, survival, metastasis, and angiogenesis. In this study, we evaluated the effects of a novel SFK inhibitor, dasatinib (BMS-354825), on SFK/FAK/p130CAS, PI3K/PTEN/Akt/mTOR, Ras/Raf/MAPK and Stats pathways in 9 HCC cell lines.
Annals of Mutagenesis is an open access, peer reviewed, scholarly journal dedicated to publish articles covering all areas of Mutagenesis.
The journal aims to promote research communications and provide a forum for doctors, researchers, physicians and healthcare professionals to find most recent advances in all areas of Mutagenesis. Annals of Mutagenesis accepts original research articles, reviews, mini reviews, case reports and rapid communication covering all aspects of mutagenesis.
Annals of Mutagenesis strongly supports the scientific up gradation and fortification in related scientific research community by enhancing access to peer reviewed scientific literary works. Austin Publishing Group brings universally peer reviewed journals under one roof thereby promoting knowledge sharing, mutual promotion of multidisciplinary science.
This study examined the effects of combined inhibition of the Src family kinases (SFK) and insulin-like growth factor 1 receptor (IGF-1R) pathways in prostate cancer (PCa) cells and in mouse models. Dual inhibition of these pathways more potently inhibited PCa tumor growth in vitro and in vivo compared to inhibiting either pathway alone. The combination treatment more completely inhibited Akt phosphorylation and downstream survival pathways in the cancer cells. This suggests that targeting both the SFK and IGF-1R pathways may be a promising therapeutic strategy for PCa by blocking independent and cooperative processes important for tumor growth and survival.
This document discusses various biomarkers used to diagnose and monitor cancer progression. It describes several common cancer antigen biomarkers like PSA, AFP, CA125, CA15-3, and CEA that are detected in serum to identify prostate, liver, ovarian, breast, and colorectal cancers respectively. Other biomarkers mentioned include hCG for germ cell tumors, thyroglobulin for thyroid cancer, heat shock proteins, glucose metabolism measured via FDG-PET scans, circulating tumor cells, and cancer stem cells. The document provides details on each biomarker's applications, typical values, and utility for cancer diagnosis and prognosis.
This document describes a bioinformatic methodology to predict synthetic lethal drug targets for cancers deficient in the tumor suppressor gene E-cadherin (CDH1). The methodology analyzes gene expression data from public databases to identify genes whose expression levels correlate with CDH1. Known synthetic lethal interactions, like between BRCA and PARP1, were correctly predicted. Several candidate synthetic lethal partners of CDH1 were identified and grouped into biological pathways. This bioinformatic approach can efficiently predict synthetic lethal targets to guide experimental validation and help develop targeted therapies for CDH1-deficient cancers.
Personalized Medicine in Diagnosis and Treatment of Cancer Maryam Rafati
The document discusses next generation sequencing (NGS) and its applications in personalized medicine for cancer diagnosis and treatment. It provides examples of several families with hereditary cancer syndromes who were analyzed using NGS to identify pathogenic variants. The results demonstrated higher response rates and prolonged progression-free and overall survival for cancer patients receiving personalized treatments based on biomarkers identified by NGS, compared to non-personalized treatments. NGS can detect somatic and germline mutations to classify cancers at a molecular level and guide precision oncology.
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Incidence of pneumonia and risk factors among patients with head and neck can...Enrique Moreno Gonzalez
This study investigated the incidence and patient- and treatment-related risk factors related to pneumonia acquired during radiotherapy (PNRT) in head and neck cancer (HNC) patients.
Gene expression analysis of a Helicobacter pyloriinfected and high-salt diet-...Enrique Moreno Gonzalez
Helicobacter pylori (H. pylori) infection and excessive salt intake are known as important risk factors for stomach cancer in humans. However, interactions of these two factors with gene expression profiles during gastric carcinogenesis remain unclear. In the present study, we investigated the global gene expression associated with stomach carcinogenesis and prognosis of human gastric cancer using a mouse model.
Acute myeloid leukemia (AML) is a hematopoietic malignancy with a dismal outcome in the majority of cases. A detailed understanding of the genetic alterations and gene expression changes that contribute to its pathogenesis is important to improve prognostication, disease monitoring, and therapy. In this context, leukemia-associated misexpression of microRNAs (miRNAs) has been studied, but no coherent picture has emerged yet, thus warranting further investigations.
Recently, a phase II clinical trial in hepatocellular carcinoma (HCC) has suggested that the combination of sorafenib and 5-fluorouracil (5-FU) is feasible and side effects are manageable. However, preclinical experimental data explaining the interaction mechanism(s) are lacking. Our objective is to investigate the anticancer efficacy and mechanism of combined sorafenib and 5-FU therapy in vitro in HCC cell lines MHCC97H and SMMC-7721.
Multicentric and multifocal versus unifocal breast cancer: differences in the...Enrique Moreno Gonzalez
This study compared the expression of E-cadherin, β-catenin, and MUC1 in multicentric/multifocal breast cancers versus unifocal breast cancers of identical tumor size and grade. The study found significantly downregulated expression of E-cadherin in multicentric/multifocal cancers compared to unifocal cancers. In contrast, no significant differences were seen in β-catenin expression between the two groups. Within the unifocal group, E-cadherin and β-catenin expression were positively correlated, but this was not seen in the multicentric/multifocal group. The results suggest multicentric/multifocal and unifocal breast cancers differ in E-
Assessment of preoperative exercise capacity in hepatocellular carcinoma pati...Enrique Moreno Gonzalez
Cardiopulmonary exercise testing measures oxygen uptake at increasing levels of work and predicts cardiopulmonary performance under conditions of stress, such as after abdominal surgery. Dynamic assessment of preoperative exercise capacity may be a useful predictor of postoperative prognosis. This study examined the relationship between preoperative exercise capacity and event-free survival in hepatocellular carcinoma (HCC) patients with chronic liver injury who underwent hepatectomy.
Overexpression of YAP 1 contributes to progressive features and poor prognosi...Enrique Moreno Gonzalez
Yes-associated protein 1 (YAP 1), the nuclear effector of the Hippo pathway, is a key regulator of organ size and a candidate human oncogene in multiple tumors. However, the expression dynamics of YAP 1 in urothelial carcinoma of the bladder (UCB) and its clinical/prognostic significance are unclear.
CXCR7 is induced by hypoxia and mediates glioma cell migration towards SDF-1a...Enrique Moreno Gonzalez
Glioblastomas, the most common and malignant brain tumors of the central nervous system, exhibit high invasive capacity, which hinders effective therapy. Therefore, intense efforts aimed at improved therapeutics are ongoing to delineate the molecular mechanisms governing glioma cell migration and invasion.
Differentiation of irradiation and cetuximab induced skin reactions in patien...Enrique Moreno Gonzalez
In order to improve the clinical outcome of patients with locally advanced squamous cell carcinoma of the head and neck (LASCCHN) not being capable to receive platinum-based chemoradiation, radiotherapy can be intensified by addition of cetuximab, a monoclonal antibody that blocks the epidermal growth factor receptor (EGFR). The radioimmunotherapy with cetuximab is a feasible treatment option showing a favourable toxicity profile. The most frequent side effect of radiotherapy is radiation dermatitis, the most common side effect of treatment with cetuximab is acneiform rash. Incidence and severity of these frequent, often overlapping and sometimes limiting skin reactions, however, are not well explored. A clinical and molecular differentiation between radiogenic skin reactions and skin reactions caused by cetuximab which may correlate with outcome, have never been described before.
Cholestasis induces reversible accumulation of periplakin in mouse liverEnrique Moreno Gonzalez
Periplakin (PPL) is a rod-shaped cytolinker protein thought to connect cellular adhesion junctional complexes to cytoskeletal filaments. PPL serves as a structural component of the cornified envelope in the skin and interacts with various types of proteins in cultured cells; its level decreases dramatically during tumorigenic progression in human epithelial tissues. Despite these intriguing observations, the physiological roles of PPL, especially in noncutaneous tissues, are still largely unknown. Because we observed a marked fluctuation of PPL expression in mouse liver in association with the bile acid receptor farnesoid X receptor (FXR) and cholestasis, we sought to characterize the role of PPL in the liver and determine its contributions to the etiology and pathogenesis of cholestasis.
Functional p53 is required for rapid restoration of daunorubicin-induced lesi...Enrique Moreno Gonzalez
This document summarizes a research article that studied the role of p53 in daunorubicin (DNR)-induced lesions in the spleen. The key findings were:
1) DNR treatment caused more rapid cell death and weight loss in the spleens of wild type mice compared to p53-null mice.
2) While wild type mouse spleens recovered normal morphology 8 days after DNR treatment, p53-null mouse spleens still had large necrotic lesions.
3) DNR treatment increased p21 levels in wild type mice but not p53-null mice, indicating p53 is required for p21 induction.
4) The results suggest p53
Post-diagnosis hemoglobin change associates with overall survival of multiple...Enrique Moreno Gonzalez
Anemia refers to low hemoglobin (Hb) level and is a risk factor of cancer patient survival. The National Comprehensive Cancer Network recently suggested that post-diagnosis Hb change, regardless of baseline Hb level, indicates the potential presence of anemia. However, there is no epidemiological study evaluating whether Hb change has direct prognostic values for cancer patients at the population level.
Cost-effectiveness of MRI for breast cancer screening in BRCA1/2 mutation car...Enrique Moreno Gonzalez
Women with mutations in BRCA1 or BRCA2 are at high risk of developing breast cancer and, in British Columbia, Canada, are offered screening with both magnetic resonance imaging (MRI) and mammography to facilitate early detection. MRI is more sensitive than mammography but is more costly and produces more false positive results. The purpose of this study was to calculate the cost-effectiveness of MRI screening for breast cancer in BRCA1/2 mutation carriers in a Canadian setting.
Impaired mitochondrial beta-oxidation in patients with chronic hepatitis C: r...Enrique Moreno Gonzalez
Hepatic steatosis is often seen in patients with chronic hepatitis C (CH-C). It is still unclear whether these patients have an impaired mitochondrial β-oxidation. In this study we assessed mitochondrial β-oxidation in CH-C patients by investigating ketogenesis during fasting.
Intraepithelial lymphocyte distribution differs between the bulb and the seco...Enrique Moreno Gonzalez
Evaluation of intraepithelial duodenal lymphocytosis (IDL) is important in celiac disease (CD). There is no established cut-off value for increased number of IELs in the bulb. We therefore investigated the relation between IEL counts in the bulb and duodenal specimens in non-celiac subjects.
Sticky siRNAs targeting survivin and cyclin B1 exert an antitumoral effect on...Enrique Moreno Gonzalez
Melanoma represents one of the most aggressive and therapeutically challenging malignancies as it often gives rise to metastases and develops resistance to classical chemotherapeutic agents. Although diverse therapies have been generated, no major improvement of the patient prognosis has been noticed. One promising alternative to the conventional therapeutic approaches currently available is the inactivation of proteins essential for survival and/or progression of melanomas by means of RNA interference. Survivin and cyclin B1, both involved in cell survival and proliferation and frequently deregulated in human cancers, are good candidate target genes for siRNA mediated therapeutics.
Association between variations in the fat mass and obesity-associated gene an...Enrique Moreno Gonzalez
It is clear that genetic variations in the fat mass and obesity-associated (FTO) gene affect body mass index and the risk of obesity. Given the mounting evidence showing a positive association between obesity and pancreatic cancer, this study aimed to investigate the relation between variants in the FTO gene, obesity and pancreatic cancer risk.
The cysteinyl leukotriene 2 receptor contributes to all-trans retinoic acid-i...Enrique Moreno Gonzalez
Cysteinyl leukotrienes (CysLTs) are potent pro-inflammatory mediators that are increased in samples from patients with inflammatory bowel diseases (IBDs). Individuals with IBDs have enhanced susceptibility to colon carcinogenesis. In colorectal cancer, the balance between the pro-mitogenic cysteinyl leukotriene 1 receptor (CysLT1R) and the differentiation-promoting cysteinyl leukotriene 2 receptor (CysLT2R) is lost. Further, our previous data indicate that patients with high CysLT1R and low CysLT2R expression have a poor prognosis. In this study, we examined whether the balance between CysLT1R and CysLT2R could be restored by treatment with the cancer chemopreventive agent all-trans retinoic acid (ATRA).
Clinical features and outcome of cryptogenic hepatocellular carcinoma compare...Enrique Moreno Gonzalez
Cryptogenic hepatocellular carcinoma (HCC) is thought to arise due to non-alcoholic fatty liver disease (NAFLD). This study investigated the prevalence, clinical features, and outcomes of cryptogenic HCC and compared them with those of HCC related to hepatitis B virus infection (HBV-HCC), hepatitis C virus infection (HCV-HCC), and alcohol (ALCHCC) in Korea.
Histololgy of Female Reproductive System.pptxAyeshaZaid1
Dive into an in-depth exploration of the histological structure of female reproductive system with this comprehensive lecture. Presented by Dr. Ayesha Irfan, Assistant Professor of Anatomy, this presentation covers the Gross anatomy and functional histology of the female reproductive organs. Ideal for students, educators, and anyone interested in medical science, this lecture provides clear explanations, detailed diagrams, and valuable insights into female reproductive system. Enhance your knowledge and understanding of this essential aspect of human biology.
8 Surprising Reasons To Meditate 40 Minutes A Day That Can Change Your Life.pptxHolistified Wellness
We’re talking about Vedic Meditation, a form of meditation that has been around for at least 5,000 years. Back then, the people who lived in the Indus Valley, now known as India and Pakistan, practised meditation as a fundamental part of daily life. This knowledge that has given us yoga and Ayurveda, was known as Veda, hence the name Vedic. And though there are some written records, the practice has been passed down verbally from generation to generation.
- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
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Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Local Advanced Lung Cancer: Artificial Intelligence, Synergetics, Complex Sys...Oleg Kshivets
Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
2. Epigenetic regulation and role of metastasis
suppressor genes in pancreatic ductal
adenocarcinoma
Wolf Arif Mardin1
Email: wolf.mardin@ukmuenster.de
Joerg Haier2*
*
Corresponding author
Email: joerg.haier@ukmuenster.de
Soeren Torge Mees1
Email: soerentorge.mees@ukmuenster.de
1
Departmwnt of General and Visceral Surgery, University Hospital of Muenster,
Albert-Schweitzer-Campus 1, 48149 Muenster, Germany
2
Comprehensive Cancer Center Muenster, University Hospital of Muenster,
Albert-Schweitzer-Campus 1, 48149 Muenster, Germany
Abstract
Background
Pancreatic ductal adenocarcinoma (PDAC) is distinguished by rapid dissemination. Thus,
genetic and/or epigenetic deregulation of metastasis suppressor genes (MSG) is a likely event
during early pancreatic carcinogenesis and a potential diagnostic marker for the disease. We
investigated 9 known MSGs for their role in the dissemination of PDAC and examined their
promoters for methylation and its use in PDAC detection.
Methods
MRNA expression of 9 MSGs was determined in 18 PDAC cell lines by quantitative RT-
PCR and promoter methylation was analyzed by Methylation Specific PCR and validated by
Bisulfite Sequencing PCR. These data were compared to the cell lines’ in vivo metastatic and
invasive potential that had been previously established. Statistical analysis was performed
with SPSS 20 using 2-tailed Spearman’s correlation with P < 0.05 being considered
significant.
Results
Complete downregulation of MSG-mRNA expression in PDAC cell lines vs. normal
pancreatic RNA occurred in only 1 of 9 investigated genes. 3 MSGs (CDH1, TIMP3 and
KiSS-1) were significantly methylated. Methylation only correlated to loss of mRNA
expression in CDH1 (P < 0.05). Bisulfite Sequencing PCR showed distinct methylation
patterns, termed constant and variable methylation, which could distinguish methylation-
3. regulated from non methylation-regulated genes. Higher MSG mRNA-expression did not
correlate to less aggressive PDAC-phenotypes (P > 0.14).
Conclusions
Genes with metastasis suppressing functions in other tumor entities did not show evidence of
assuming the same role in PDAC. Inactivation of MSGs by promoter methylation was an
infrequent event and unsuitable as a diagnostic marker of PDAC. A distinct methylation
pattern was identified, that resulted in reduced mRNA expression in all cases. Thus, constant
methylation patterns could predict regulatory significance of a promoter’s methylation prior
to expression analysis and hence present an additional tool during target gene selection.
Keywords
Pancreatic ductal adenocarcinoma, PDAC, Metastasis suppressor gene, Methylation,
Epigenetics, Promoter
Background
Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy with a dismal
5-year survival rate of less than 5%, a median survival of 6 months and a mortality to
incidence ratio of 0.98 [1,2]. While surgical intervention represents the only curative
treatment option, more than 80% of patients with PDAC present with irresectable disease
[3,4]. Yet even after surgery, 5-year survival in specialized centers reaches no more than 25%
with (neo)adjuvant treatment and remains below 15% without it [5,6]. Existing strategies
seem to be exhausted and novel markers as well as new therapeutic targets are needed to both
move forward the time of diagnosis and increase treatment efficacy.
Genetic alterations to factors and pathways underlying pancreatic ductal carcinogenesis, such
as KRAS, TP53, SMAD4 and Hedgehog are under close investigation [7]. However, beside
genetic alterations, epigenetic mechanisms for gene inactivation, such as transcriptional
silencing by promoter methylation, seem to be important in the pathogenesis of PDAC [8].
Recent studies have shown that aberrant methylation of CpG-islands is a common mechanism
associated with the silencing of tumor-suppressor and cancer-related genes in pancreatic
cancers [9], among them BRCA1, APC, and p16INK4a [10-12]. These aberrant methylation
patterns and the resulting changes in gene expression may over the long term be drawn upon
as therapeutic targets. A more tangible use however may be their exploit as biomarkers; these
would ideally enable earlier diagnosis or even screening for pancreatic cancer. Presently, the
disease is mostly diagnosed by the onset of clinical symptoms which occur at later stages,
when a curative approach, i.e. resection, is mostly no longer possible. Moving forward the
time of diagnosis would thus increase survival through identification at more treatable disease
stages [13,14]. As a consequence, biological changes used for the detection of PDAC should
ideally be present early on in pancreatic carcinogenesis and precancerous lesions. Prevalence
of low-level aberrant methylation has indeed been detected in well-defined pancreatic
intraductal neoplasia (PanIN) lesions and overexpression of DNA methyltransferase 1 has
been found in precancerous conditions and it increased with disease progression [15].
Detection of aberrant methylation in body fluids such as serum, urine, pancreatic juice or
sputum could also be a useful marker for PDAC [16]. For example, analysis of NPTX2
[17,18], RASSF1A [19], cyclin D2 [20], ppENK and pi6 [21] hypermethylation may enable
4. the creation of a highly sensitive and less invasive panel of markers for pancreatic cancer.
These findings raise hopes that early markers of PDAC may be developed based on aberrant
methylation patterns.
Due to the disease’s rapid progression and early metastasis formation, metastasis suppressor
genes (MSGs) may be deregulated early during pancreatic carcinogenesis. Clinically and
experimentally, primary tumor development and metastasis formation are distinct processes.
Metastasis suppressors are defined as inhibitors of metastasis at any step of the metastatic
cascade without interfering with primary tumor growth [22]. The discovery of these
endogenous molecules that exclusively inhibit metastasis and the understanding of their
actions suggest that metastasis is an amenable therapeutic target. However, only few of the
currently known MSGs have been investigated in pancreatic cancer so far. In the present
study, we selected a comprehensive panel of MSGs based on review of metastasis
suppressors by Shevde and Welch [23].
Taken together, our review of the current literature revealed 1) earlier diagnosis of PDAC
would increase survival, 2) hypermethylation occurs early during pancreatic carcinogenesis
and 3) metastasis suppressor genes have not been sufficiently researched in this context.
Accordingly, we investigated a number of known MSGs for their influence on PDAC
dissemination and the methylation status of their promoter region In order to identify possible
PDAC biomarker candidates.
Methods
Ethics statement
All animal experiments were conducted in accordance with the national guidelines for the
care and use of laboratory animals, and the experimental protocol was approved by the state
agency for animal welfare of North Rhine-Westphalia (LANUV; NRW, Germany).
Genes of interest
Analyses were carried out for 9 known MSGs, as reviewed by Shevde and Welch [23]:
BRMS1, CD82, CDH1, KiSS-1, MAP2K4, MED23, NDRG1, TIMP3 and TXNIP.
Cell lines
18 human PDAC cell lines were analyzed in vitro. All cell lines were obtained from
American Type Culture Collection, except A818, PT45, HPAF2 and MiaPaCa2 which were
originally obtained from American Type Culture Collection and provided by H. Kalthoff,
Department of Surgery, University Hospital Kiel, Germany and H. Hotz, Department of
Surgery, Charite CBF, University Hospital Berlin, Germany. Cells were maintained in
recommended growth media, and all media were supplemented with 10% heat inactivated
fetal bovine serum (Gibco / Invitrogen, Karlsruhe, Germany) and were mycoplasma negative.
For culturing, they were incubated at 37°C in humidified air with 5% or 10% CO2. The
medium was replaced twice a week, and cells were maintained by serial passaging after
trypsinization with 0.1% trypsin.
5. Animal model and biological classification of cell lines
An orthotopic implantation tumor model using four-week-old male nude mice (Crl:NU/NU-
nuBR) had been carried out previously [24]. In brief, four-week-old male nude mice
(Crl:NU/NU-nuBR) were injected subcutaneously with each human PDAC cell line. These
mice were euthanized after 3 to 4 weeks and the donor tumors collected, fragments of which
were inserted orthotopically into another mouse’s pancreatic parenchyma (n = 10 mice per
cell line). HS766T and PL45 failed to form tumors in donor mice and thus were excluded
from in vivo, but not from in vitro analysis. After a growth phase of 12 weeks, primary tumor
volume, local infiltration and patterns of local and systemic metastases were assessed
systematically as previously described [25]. The results were compiled into a score each for
metastasis and invasion.
Nucleic acid preparation
DNA and RNA extraction from cell lines was performed using the DNeasy Blood & Tissue
and the RNeasy Mini Kit from Qiagen (Hildesheim, Germany) according to the
manufacturer’s specifications. A total RNA preparation from human pancreas was acquired
from Applied Biosystems (Darmstadt, Germany). RNA samples were stored at −80°C.
CDNA was generated using the High Capacity cDNA Reverse Transcription Kit from
Applied Biosystems according to the manufacturer’s specifications. CDNA was stored at
−20°C. Purity and concentration of nucleic acids were measured in a biophotometer
(Eppendorf, Hamburg, Germany).
Bisulfite modification
Bisulfite modification of DNA was performed with EpiTect Bisulfite Kits from Qiagen
according to the manufacturer’s specifications. Obtained products were stored at −20°C.
Methylation assays
Methylation specific PCR (MSP) and Bisulfite sequencing PCR (BSP) were carried out as
previously described for in vitro cell lines [26]. BSP results were analyzed with the Beckman
Coulter CEQ 8800 Genetic Analysis System software v9.0 using C- to T-peak ratios to define
a CpG-dinucleotide as methylated, unmethylated or heterogeneously methylated for each
CpG-dinucleotide. Point values were assigned to each CpG-dinucleotide according to its
methylation status as follows: unmethylated: 0; heterogeneously methylated: 1; methylated:
2. A methylation-score was calculated for each gene in each cell line by using the average
point value of all investigated CpG-dinucleotides for that gene, resulting in values from 0.0
(completely unmethylated) to 2.0 (completely methylated).
Quantitative reverse transcriptase-PCR
Quantitative reverse transcriptase-PCR (qRT-PCR) had been previously performed for in
vitro cell lines [26]. Calculations were carried out using the qBase algorithm in Microsoft
Excel using the 2-∆∆CT
Method with PPIB and HPRT1 as reference genes [27].
6. Statistical analysis
Statistical analysis was performed using 2-tailed Spearman’s correlation. 95% confidence
intervals were calculated and P < 0.05 was considered significant. SPSS 20.0 (SPSS Inc.,
Chicago, IL, USA) was used for these calculations.
Results
Animal model: tumor biology of the PDAC cell lines
In vivo metastatic potential had been previously investigated for 16 PDAC cell lines and
quantified by a metastasis- and invasion-score [24]. In brief, the scores were calculated by
crediting one point for every local infiltration, every colonized organ, and for multiple
metastatic lesions per organ. Score values represent mean sums of the obtained credit points
for all mice in a group .
Metastasis suppressor gene mRNA expression
We investigated the expression of 9 MSGs by qRT-PCR for 18 PDAC cell lines and normal
human pancreatic RNA. Gene selection was performed at the beginning of the investigation
and reflects the known MSGs at that time. In 6/9 genes (BRMS1, CD82, CDH1, MED23,
NDRG1 and TXNIP) and both reference genes (HPRT1, PPIB) mRNA was detected in all
samples. Of the remaining 3 genes, KiSS-1 was detected in all cell lines except MiaPaCa2;
MAP2K4 was detected in all cell lines except A818, MPanc 96, PaTu 8902, PaTu 8988S and
PaTu 8988 T; TIMP3 was detected in all cell lines except A818, MiaPaCa2, PaTu 8902 and
PaTu 8988 T.
Of note: Isolation of in vivo samples of sufficient quality (RIN > 7.0; tumor cells >80% in
microscopic analysis) was not possible. The underlying reasons are presented in the
discussion section.MAP2K4, TIMP3 and TXNIP were downregulated in most PDAC cell
lines while BRMS1 and KiSS-1 showed predominant upregulation vs. normal pancreatic
RNA (Figure 1 and [26]).
Figure 1 Metastasis suppressor gene mRNA expression. Legend: MRNA expression
(qRT-PCR) of metastasis suppressor genes in PDAC cell lines and normal pancreatic RNA as
previously determined by our group [26]. Normal pancreatic RNA is highlighted,
demonstrating that downregulation of metastasis suppressor genes in PDAC cell lines vs.
normal RNA was not a uniform occurrence.
Methylation specific PCR
MSP analysis of CpG-islands in the promoter region of the investigated genes showed 3
patterns. 1) and 2): Only one band was found per MSP: either the PCR with the methylation-
specific primer showed a band while the PCR with the unmethylation-specific primer did not
or vice versa. 3) Both PCRs from the same MSP showed bands simultaneously. The results
were therefore classified as homogenously methylated, homogenously unmethylated or
heterogeneously methylated (Figure 2a). No gene was methylated in all cell lines while
several genes were unmethylated across all cell lines (Figure 2b). Gel pictures are provided in
Additional file 1.
7. Figure 2 Methylation specific PCR. Legend: A) Example of a methylation specific PCR:
KiSS-1. Samples are noted above the image and primer specificity is declared as U, specific
for unmethylated template, or M, specific for methylated template. 3 distinct signal patterns
were observed: either only the U- or the M-band was present, or both bands were present.
Cell lines were thus identified as unmethylated (e.g. MPanc96), methylated (e.g. PaTu8988S)
or heterogeneously methylated (e.g. CAPAN2). B) MSP-analysis for all cell lines and genes.
Each column represents a gene and its methylation status in a cell line is indicated by
partially filled circles. Methylation status for each cell line was determined as illustrated in
Figure 3A for the example for KiSS-1. *different results for MSP- vs. BSP-analysis (compare
to Figure 4).
Bisulfite sequencing PCR and direct sequencing
Three types of signals were present at the CpG-sites: 1) A T-peak and no C-peak; classified
as an unmethylated CpG-dinucleotide 2) A C-peak and no T-peak; classified as a methylated
CpG-dinucleotide 3) Superimposing C- and T-peaks; classified as heterogeneously
methylated CpG-dinucleotide. 5 of the 9 investigated genes (CD82, MAP2K4, MED23,
NDRG1, TXNIP) showed no methylation throughout the entire sequenced region (Figure 3).
The remaining 4 genes, BRMS1, CDH1, KiSS-1 and TIMP3 showed significant methylation,
with differing methylation patterns and intensities (Figure 4.).
Figure 3 Overview of bisulfite sequencing methylation scores. Legend: Increasingly
colored in circles represent methylation score ranges from bisulfite sequencing. Residual
methylation was present in BRMS1, which however still translated into an overall
methylation score of <0.4. Detailed methylation maps are provided in Figure 5.
Figure 4 Methylation volatility. Legend: Each box represents one sequenced CpG in 5′ to 3′
order whose methylation status is indicated by coloration. Some cell lines have uniform
methylation patterns throughout the entire investigated region of a gene (e.g. CDH1 in
MiaPaCa2: constant methylation pattern), while a more inconsistent pattern is present in
other cases (e.g. PL45 in TIMP3: variable methylation pattern).
Correlation between MSP and BSP-direct sequencing
MSP-status and the BSP methylation-score were highly correlated in the methylated genes (P
< 0.005, except TIMP3: P < 0.05). However, differing results between MSP and BSP-direct
sequencing were discovered for 2 cell lines in TIMP3 and 11 cell lines in KiSS-1. In all of
these instances, BSP-direct sequencing was more sensitive to methylation than MSP (Figure
2b and Figure 3). The unmethylated genes were identified as such by both methods with only
BRMS1 showing minor methylation in BSP-direct sequencing.
Overall, BSP proved more sensitive to methylation than MSP, delivering an explanation for
the only slightly differing results in KiSS-1. In TIMP3 however, greater discrepancies were
encountered, suggesting the possibility of differential methylation patterns within the
promoter island as the BSP-sequenced region did not include the region in which the MSP
primers were situated.
8. Methylation patterns
We identified two distinct methylation patterns in the four genes with significant promoter
methylation. The first pattern was dubbed variable methylation: In BRMS-1, KiSS-1 and
TIMP3, volatility of methylation was high with parallel occurrence of methylated,
heterogeneously methylated and unmethylated CpGs at a single methylation site (Figure 4).
The second pattern, present in CDH1, was termed constant methylation: CDH1 was only
methylated in 2/18 (11%) cell lines. In contrast to the methylation-variability observed in the
first group, the CDH1 promoter had a constant methylation pattern for single cell lines: the
cell lines displayed either uniformly high or uniformly low methylation across the entire
sequenced promoter region with low methylation volatility. Promoter methylation and
mRNA expression data from 2 additional metastasis suppressors, AKAP12 and SERPINB5,
were included for methylation pattern analysis. Our group has previously investigated these
genes and identified a stable methylation pattern for the SERPINB5 promoter while AKAP12
displayed variable methylation [26]. Figure 5A is a graphic representation of the differences
in overall methylation intensity and variability of methylation intensity. Differences in the
standard deviation of the methylation scores was used as a mathematical basis for the
definition of variable or constant methylation patterns: methylation score standard deviation
of ≤0.09 was defined as constant methylation, and >0.09 as variable methylation (Figure 5F,
B).
Figure 5 Methylation volatility. Legend: A) Each row represents the sequenced CpG sites
of one gene with each bar standing for one CpG-site in 5′ to 3′ direction. Bar height denotes
the average methylation-score of that CpG-site across all cell lines. Variably methylated
genes (AKAP12, BRMS1, TIMP3, KiSS-1) showed generally higher methylation with greater
volatility than genes with constant methylation (CDH1, SERPINB5). B) The observation in A
is quantified by differing levels of methylation volatility: genes with constant methylation
had lower standard deviation of methylation levels than those with variable methylation.
In summary, we identified three types of promoter methylation: 1) Genes with completely
unmethylated promoters 2) Genes whose promoters were methylated in most cell lines and
that showed highly variable methylation intensity (methylation score standard deviation
>0.09), dubbed variable methylation. 3) Gene promoters that were methylated in few cell
lines and with constant methylation intensity (methylation score standard deviation ≤0.09),
dubbed constant methylation.
Correlation between promoter methylation and loss of mRNA expression
The expression levels of the 4 methylated genes were investigated for differential mRNA
expression in correlation to their promoter methylation status. The priorly investigated genes,
SERPINB5 (constant methylation pattern) and AKAP12 (variable methylation pattern) were
included for this analysis. In the genes with constant methylation (CDH1 and SERPINB5) the
significantly methylated cell lines had the lowest mRNA expression across all cell lines. For
the variably methylated genes (AKAP12, BRMS1, KiSS-1 and TIMP3), no significant
correlations between methylation-score and mRNA expression were detected. Thus, constant
methylation patterns exerted influence on mRNA expression in all cases, while all instances
of variable methylation patterns had no influence on mRNA expression (Table 1).
9. Table 1 Correlation of methylation patterns to mRNA expression
Constant methylation Variable methylation
CDH1 SERPINB5 AKAP12 BRMS1 KiSS-1 TIMP3
Methylation score vs. mRNA expression P < 0.05 P < 0.05 n.s. n.s. n.s. n.s.
Legend: Constant methylation patterns are correlated to loss of mRNA expression, while
variable methylation patterns are not.
Correlation between metastasis suppressor gene mRNA expression and tumor
biology
No significant correlations between MSG mRNA expression and tumor biology were
detected. Both invasion and metastasis scores were not correlated to expression of the nine
investigated metastasis suppressor genes (Additional file 2: Table S1).
Discussion
Although aberrant methylation is a frequent epigenetic event for a number of genes in
pancreatic cancer, in the present study, promoter methylation was present in only 4 of 9
investigated genes and only resulted in reduced mRNA expression one of them, CDH1.
For further analysis of methylation levels, we included 2 previously investigated, methylated
genes: AKAP12 and SERPINB5, increasing the number of methylated genes to 6. For this
group of genes we found 2 distinct and mutually exclusive types of methylation, either
“constant” or “variable” methylation. Variably methylated genes showed considerable
methylation volatility for single CpG sites with methylation levels changing from completely
unmethylated to completely methylated several times throughout the sequenced promoter
region. The genes in this group were AKAP12, BRMS1, KiSS-1 and TIMP3 -- genes for
which there was no correlation between promoter methylation and mRNA expression. Thus,
variable methylation, even though often occurring at high levels, did not translate into loss of
mRNA expression. This suggests that variable methylation patterns could be a by-product of
tumor dedifferentiation and possibly DNMT-1 (DNA methyltransferase 1) overexpression
without regulatory links to mRNA-expression. DNMT1 protein expression has been reported
to increase progressively during the stages of pancreatic carcinogenesis and was found to be
associated with tumor aggressiveness, suggesting that protein overexpression of DNMT1 and
ensuing hypermethylation may be involved in multistage pancreatic carcinogenesis -- even
though it may not translate into loss of mRNA expression [15]. In contrast, for SERPINB5
and CDH1, the genes with constant methylation patterns, the methylated cell lines showed
the lowest mRNA expression across all investigated samples, correlating this pattern of
methylation to loss of mRNA expression. These observations suggest that the functional
relevance of promoter methylation can be predicted even before carrying out quantitative
reverse transcriptase PCR, based only on methylation patterns. While confirmation by qRT-
PCR will still be necessary, target selection could be made more efficient through the pre-
selection of genes of interest according to their methylation pattern.
Surprisingly, overexpression of some investigated MSGs was found in many tumor cell lines
compared to normal pancreatic RNA. For example, BRMS1 demonstrated nearly uniform
upregulation in the PDAC cell lines (94%, 17/18 cell lines). This gene was first described in
breast cancer and was more recently found to decrease metastatic potential in that tumor
entity by upregulating microRNA miR-146 [28] . While we do not know whether miR-146
10. induction by BRMS1 also occurs in PDAC, it is known that miR-146 is overexpressed in
pancreatic cancer vs. both normal pancreas and pancreatitis, suggesting that miR-146 has an
oncogenic role in pancreatic cancer [29]. This corresponds to our present findings of nearly
uniform upregulation of BRMS1 in PDAC vs. normal tissue. Further investigation of the
BRMS1 / miR-146 axis in PDAC appears to be warranted.
We encountered substantial difficulties regarding mRNA expression analysis from in vivo
tissues: The recovery of mRNA samples of sufficient quality (RIN > 7.0; tumor cells >80% in
microscopic analysis) was not possible due to 2 main reasons: 1) The orthotopic tumor
specimens decayed too quickly, likely due to autodigestion before complete permeating of
the RNAse inactivating agent, resulting in insufficient RIN values <7.0. 2) The metastases
were simply too small for recovery of sufficient sample volume. In addition, it was not
possible to control these samples for contamination by normal cells without destroying the
sample. Thus, we proceeded without these analyses as they were technically unfeasible.
None of the investigated genes showed any correlation to the metastatic or infiltrative
potential of 16 PDAC cell lines in vivo. Instead, some were significantly upregulated in the
PDAC cell lines vs. normal pancreatic RNA. The present results suggest that most known
metastasis suppressor genes do not assume their ascribed roles in PDAC and that some
metastasis suppressors even undergo overexpression in this tumor entity. Further, promoter
methylation does not seem to be a prominent mechanism of loss of expression for metastasis
suppressor genes in PDAC as most genes’ promoter islands showed no methylation at all.
Even when methylation did occur, it did not result in loss of mRNA in most cases. Thus,
assays for metastasis suppressor gene promoter methylation do not appear to be suitable tools
for the detection of PDAC. However, when methylation did induce loss of mRNA
expression, a distinct methylation pattern was observed in each case, termed constant
methylation. To our knowledge, methylation patterns have not yet been correlated to
translation into loss of mRNA expression. Yet methylation patterns may indeed be able to
predict whether or not loss of expression occurs when methylation is present. After a first
global methylation analysis, information on methylation patterns could help to select genes
for further, i.e. functional, analysis. Detailed knowledge of epigenetic alterations and
associated molecular mechanisms during pancreatic tumorigenesis and metastasis will
broaden the biological understanding of the disease and help devise novel targets for earlier
diagnosis and increasingly effective therapies.
Conclusions
In summary, our results suggest that metastasis suppressor genes may play different roles in
PDAC compared to other tumor entities, a hypothesis that must be confirmed by in depth
analysis of single genes. In addition, we describe promoter methylation patterns that were
correlated to functional relevance: Patterns of constant promoter methylation appear to
predict methylation-associated control of gene expression.
Competing interests
All authors declare that they have no competing interests.
11. Authors’ contributions
Study was conceived by JH and WAM. Experiments were carried out by STM and WAM.
Statistical analysis was carried out by WAM, STM and JH. Manuscript was written by JH,
STM and WAM. All authors read and approved the final manuscript.
Acknowledgements
The authors thank F. Spiecker and C. Kemming for their support and expert technical
assistance and A. Enns for his critical input and assistance with experimental design. The
authors also thank H. Kalthoff (Department of Surgery, University Hospital Kiel, Germany)
for the cell lines A818 and PT45, and H. and B. Hotz (Department of Surgery, Charite CBF,
University Hospital Berlin, Germany) for the cell lines HPAF-2 and MiaPaCa2, as well as
their advice with the murine model. Quantitative Real Time PCR was performed at the
Integrated Functional Genomics (IFG), Core Unit of the Interdisciplinary Center for Clinical
Research (IZKF) Muenster, Germany.
Grants
This study was supported by a grant from the Foerderverein Peter Geiger e.V., Beilstein,
Germany and a grant from Innovative Medizinische Forschung (IMF), Muenster, Germany.
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Additional files
Additional_file_1 as JPEG
Additional file 1 MSP Gels.
Additional_file_2 as DOC
Additional file 2: Table S1 Correlation of metastasis suppressor expression to tumor
biology. Legend: No significant correlations between metastasis suppressor gene expression
and in vivo tumor biology were identified.