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.
A new assay for measuring chromosome instability (CIN) and identification of...Enrique Moreno Gonzalez
Aneuploidy is a feature of most cancer cells that is often accompanied by an elevated rate of chromosome mis-segregation termed chromosome instability (CIN). While CIN can act as a driver of cancer genome evolution and tumor progression, recent findings point to the existence of a threshold level beyond which CIN becomes a barrier to tumor growth and therefore can be exploited therapeutically. Drugs known to increase CIN beyond the therapeutic threshold are currently few in number, and the clinical promise of targeting the
CIN phenotype warrants new screening efforts. However, none of the existing methods, including the in vitro micronuclei (MNi) assay, developed to quantify CIN, is entirely satisfactory.
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.
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.
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.
Search for atoxic cereals: a single blind, cross-over study on the safety of...Enrique Moreno Gonzalez
Cereals of baking quality with absent or reduced toxicity are actively sought as alternative therapy to a gluten-free diet (GFD) for patients with coeliac disease (CD). Triticum monococcum, an ancient wheat, is a potential candidate having no toxicity in in-vitro and exvivo studies. The aim of our study was to investigate on the safety of administration of a single dose of gluten of Tm in patients with CD on GFD.
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.
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.
A new assay for measuring chromosome instability (CIN) and identification of...Enrique Moreno Gonzalez
Aneuploidy is a feature of most cancer cells that is often accompanied by an elevated rate of chromosome mis-segregation termed chromosome instability (CIN). While CIN can act as a driver of cancer genome evolution and tumor progression, recent findings point to the existence of a threshold level beyond which CIN becomes a barrier to tumor growth and therefore can be exploited therapeutically. Drugs known to increase CIN beyond the therapeutic threshold are currently few in number, and the clinical promise of targeting the
CIN phenotype warrants new screening efforts. However, none of the existing methods, including the in vitro micronuclei (MNi) assay, developed to quantify CIN, is entirely satisfactory.
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.
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.
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.
Search for atoxic cereals: a single blind, cross-over study on the safety of...Enrique Moreno Gonzalez
Cereals of baking quality with absent or reduced toxicity are actively sought as alternative therapy to a gluten-free diet (GFD) for patients with coeliac disease (CD). Triticum monococcum, an ancient wheat, is a potential candidate having no toxicity in in-vitro and exvivo studies. The aim of our study was to investigate on the safety of administration of a single dose of gluten of Tm in patients with CD on GFD.
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.
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.
Functional p53 is required for rapid restoration of daunorubicin-induced lesi...Enrique Moreno Gonzalez
The tumour suppressor and transcription factor p53 is a major determinant of the therapeutic response to anthracyclines. In healthy tissue, p53 is also considered pivotal for side effects of anthracycline treatment such as lesions in haematopoietic tissues like the spleen. We used a Trp53null mouse to explore the significance of p53 in anthracycline (daunorubicin) induced lesions in the spleen.
Functional p53 is required for rapid restoration of daunorubicin-induced lesi...Enrique Moreno Gonzalez
The tumour suppressor and transcription factor p53 is a major determinant of the therapeutic response to anthracyclines. In healthy tissue, p53 is also considered pivotal for side effects of anthracycline treatment such as lesions in haematopoietic tissues like the spleen. We used a Trp53null mouse to explore the significance of p53 in anthracycline (daunorubicin) induced lesions in the spleen.
Epigenetic regulation and role of metastasis suppressor genes in pancreatic d...Enrique Moreno Gonzalez
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.
Abstract
Aberrant mucin-type O-glycosylation by glycosyltransferases is a well-described hallmark of many cancers and is also associated with additional non-cancerous developmental and metabolic disorders. The current review focuses on N-acetylgalactosaminyltransferase genes (GALNT) and proteins (GalNAcTs) to illustrate their importance in cancer biology. Aberrant O-glycosylation by GalNAcTs activates a wide range of proteins that carry out interactions of sessile and motile cells affecting organogenesis, responses to agonists and stimulating hyperproliferation and metastatisation of neoplastic cells. As genome-wide analyses have provided abundant clues regarding under- or over-expressed genes that characterize different types of cancers, GALNTs and their transferase products have attracted attention by being unexpected actors in neoplastic contexts. We intend to review the current knowledge on GALNTs and their encoded transferases in cancer and suggest what could be the significance of such information in cancer pathogenesis and management.
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.
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.
Estimation of G6pd Status in the Rajbangshi Population of SushrutanagarIOSR Journals
The Glucose 6 Phosphate Dehydrogenase (G6PD) deficiency renders the cells susceptible to severe hemolysis under oxidative stress producing conditions. Due to considerable genetic heterogeneity it is found to be distributed in an unequal manner in different parts of the world including India. The present study was undertaken to find the distribution of G6PD deficiency in the Rajbangshi population group of Sushrutanagar, Darjeeling district. The Rajbangshis are one of the oldest tribes residing in North Bengal. A quantitative assay of the G6PD activity was performed in the Rajbangshi population and the non Rajbangshi control population of the same area. The G6PD deficiency was found in 12 percent of the Rajbangshi population studied which is significantly higher than the value of 3.3 percent found in the controls with the lower and upper confidence limit for the population Odds ratio being 3.58 and 25.93 respectively at 95% confidence interval. This high rate of G6PD deficiency in the population group studied poses them to a greater risk for several oxidative stress producing conditions including some commonly used antimalarial drug like Primaquin
This presentation (in English) made at ONCOTRANS in Besançon on Friday 3rd 2017 reviews the potential for TGF-beta inhibition in hepatocellular carcinoma based on preclinical and clinical data.
The Single Nucleotide Polymorphisms (SNPS) of Vascular Endothelial Growth Fac...Agriculture Journal IJOEAR
Abstract—
Aim: The aim of the present work was to evaluate associations between the risk of endometriosis and -460C/T (rs833061) and +405G/C (rs2010963) polymorphisms in the VEGF gene.
Methodology: In the present study, we examined group of 100 patients with endometriosis and 100 controls. Genomic DNA was extracted from peripheral blood. Determination of genes polymorphic variants was made using polymerase chain reaction-restriction fragment length polymorphism technique (PCR-RFLP).
Results: Presented study showed statistically significant increase in the endometriosis development risk for the -460T/T genotype (OR 3.39; 95% CI, 1.60-7.13; p = 0.002) and for the -460T allele (OR 2.49; 95% CI, 1.64-3.78; p <.0001), as well as for the +405C/C genotype (OR 2.16; 95% CI, 1.047-4.48; p = 0.035) in patients with endometriosis in comparison with healthy control group. We also observed positive association of the +405C/C genotype (OR 0.26; 95% CI, 0.08-0.79; p = 0.019) as well as the +405C allele occurrence with an increased endometriosis development risk (OR 0.31; 95% CI, 0.19-0.71; p = 0.005), assessed by the degree of rASRM classification stages.
Conclusion: The results support the hypothesis that the -460C/T and +405G/C polymorphisms of the VEGF gene may be associated with endometriosis occurrence in Poland.
Keywords— endometriosis, genetic polymorphisms, VEGF.
hMSH2 Gly322Asp (rs4987188) Single nucleotide polymorphism and the risk of br...Agriculture Journal IJOEAR
Aim: Breast cancer is the most common cancer in women both in the developed and less developed world. The reported study was designed to explore associations between hMSH2 - Gly322Asp (1032G>A, rs4987188) single nucleotide polymorphism (SNP) and the risk of breast carcinoma in the Polish women.
Material and methods: Blood samples were obtained from women with breast cancer (n=225), treated at the Department of Oncological Surgery and Breast Diseases, Polish Mother’s Memorial Hospital – Research Institute between the years 2005 and 2012. A control group included 220 cancer-free women. Genomic DNA was isolated and the SNP Gly322Asp of hMSH2 was determined by High-Resolution Melter method. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for each genotype and allele.
Results: This study revealed that single nucleotide polymorphism Gly322Asp of hMSH2 is associated with both breast cancer risk and grading. Moreover, it can be linked with breast carcinoma tumor size and lymph node status. The Asp allele in patients may be a risk factor for breast carcinoma (OR 5.12; 95% CI 3.77 –6.97, p<.0001).
Conclusions: Gly322Asp single nucleotide polymorphism of hMSH2 may be a risk factor of breast cancer in the Polish women.
Similar to Variant G6PD levels promote tumor cell proliferation or apoptosis via the STAT3/5 pathway in the human melanoma xenograft mouse mode (20)
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.
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.
Multicentric and multifocal versus unifocal breast cancer: differences in the...Enrique Moreno Gonzalez
The aim of this study was to evaluate the expression of the cell adhesion-related glycoproteins MUC-1, β-catenin and E-cadherin in multicentric/multifocal breast cancer in comparison to unifocal disease in order to identify potential differences in the biology of these tumor types.
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.
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.
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.
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.
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
HOT NEW PRODUCT! BIG SALES FAST SHIPPING NOW FROM CHINA!! EU KU DB BK substit...GL Anaacs
Contact us if you are interested:
Email / Skype : kefaya1771@gmail.com
Threema: PXHY5PDH
New BATCH Ku !!! MUCH IN DEMAND FAST SALE EVERY BATCH HAPPY GOOD EFFECT BIG BATCH !
Contact me on Threema or skype to start big business!!
Hot-sale products:
NEW HOT EUTYLONE WHITE CRYSTAL!!
5cl-adba precursor (semi finished )
5cl-adba raw materials
ADBB precursor (semi finished )
ADBB raw materials
APVP powder
5fadb/4f-adb
Jwh018 / Jwh210
Eutylone crystal
Protonitazene (hydrochloride) CAS: 119276-01-6
Flubrotizolam CAS: 57801-95-3
Metonitazene CAS: 14680-51-4
Payment terms: Western Union,MoneyGram,Bitcoin or USDT.
Deliver Time: Usually 7-15days
Shipping method: FedEx, TNT, DHL,UPS etc.Our deliveries are 100% safe, fast, reliable and discreet.
Samples will be sent for your evaluation!If you are interested in, please contact me, let's talk details.
We specializes in exporting high quality Research chemical, medical intermediate, Pharmaceutical chemicals and so on. Products are exported to USA, Canada, France, Korea, Japan,Russia, Southeast Asia and other countries.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
2. Variant G6PD levels promote tumor cell
proliferation or apoptosis via the STAT3/5 pathway
in the human melanoma xenograft mouse modelA
Tao Hu1,2,†
Email: hutaoxiyang_1012@126.com
Chunhua Zhang1,3,†
Email: kmc200401111@yahoo.com.cn
Qiongling Tang1,4,†
Email: Qiongli420@163.com
Yanan Su5
Email: gpmyas@genpromarkers.com
Bo Li6
Email: boli1212@gmail.com
Long Chen1
Email: lonechen1983@hotmail.com
Zheng Zhang1
Email: Zhangzhkmu@163.com
Tianchi Cai1
Email: ctianchikmu@yahoo.com
Yuechun Zhu1*
*
Corresponding author
Email: Zhuyuechun20091119@126.com
1
Department of Biochemistry and Molecular Biology, Kunming Medical
University, Kunming 650031, China
2
Department of laboratory medicine, The Third People’s Hospital of Yunnan
Province, Kunming, China
3
Yunnan Provincial Maternal and Child Health Hospital, Kunming, China
4
Shenzhen Luohu Maternal and Child Health Hospital, Shenzhen, China
5
GenProMarkers, Inc., 9700 Great Seneca Highway, Suite182, Rockville, MD
20850, USA
6
Department of laboratory animal, Kunming Medical University, Kunming,
China
3. †
Equal contributors.
Abstract
Background
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.
Methods
HEM (human epidermal melanocyte) cells and human melanoma cells with the wild-type
G6PD gene (A375-WT), G6PD deficiency (A375-G6PD∆), G6PD cDNA overexpression
(A375-G6PD∆-G6PD-WT), and mutant G6PD cDNA (A375-G6PD∆-G6PD-G487A) were
subcutaneously injected into 5 groups of nude mice. Expressions of G6PD, STAT3, STAT5,
cell cycle-related proteins, and apoptotic proteins as well as mechanistic exploration of
STAT3/STAT5 were determined by quantitative real-time PCR (qRT-PCR),
immunohistochemistry and western blot.
Results
Delayed formation and slowed growth were apparent in A375-G6PD∆ cells, compared to
A375-WT cells. Significantly decreased G6PD expression and activity were observed in
tumor tissues induced by A375-G6PD∆, along with down-regulated cell cycle proteins cyclin
D1, cyclin E, p53, and S100A4. Apoptosis-inhibited factors Bcl-2 and Bcl-xl were up-
regulated; however, apoptosis factor Fas was down-regulated, compared to A375-WT cells.
Moderate protein expressions were observed in A375-G6PD∆-G6PD-WT and A375-G6PD∆-
G6PD-G487A cells.
Conclusions
G6PD may regulate apoptosis and expression of cell cycle-related proteins through
phosphorylation of transcription factors STAT3 and STAT5, thus mediating formation and
growth of human melanoma cells. Further study will, however, be required to determine
potential clinical applications.
Keywords
Glucose-6-phosphate dehydrogenase, Melanoma, Nude mice, Apoptosis, Signal transduction
and transcription activator
Background
Glucose-6-phosphate dehydrogenase (G6PD) is a critical enzyme in mammalian erythrocytes
that catalyzes the first reaction in the pentose-phosphate pathway [1]. G6PD abnormalities
affect as much as 7.5% of the global population, with a wide range of occurrence based on
4. geographic distribution. These abnormalities occur in as little of 0.1% of certain Japanese
populations to more than 35% in some African and European populations [2]. In fact, G6PD
was the first prevalent enzyme deficiency to be characterized by the World Health
Organization (WHO) in 1984 [3]. Around the world, between 140 and 160 distinct G6PD
mutations have been reported [1].
In China, at least 24 mutations have been detected in the G6PD gene [4,5]. Our previous
study showed that G6PD Mahidol (487G>A) was the most common G6PD variant in the
Achang ethnic group of Yunnan Province [6]. Otherwise, G6PD Mahidol is a common
deficient variant caused by a (163)glycine-serine mutation that occurs in about 15% of
individuals in populations across Southeast Asia [7,8]. The prevalence of this mutation can be
accounted for by strong positive selection over the past 1500 years that occurred in response
to certain parasites, including malaria-causing agents such as Plasmodium vivax and
Plasmodium falciparum, that target humans [9]. Like many mutations involved in parasitic
resistance, G6PD Mahidol is a relatively recent polymorphism generally referred to as a
‘loss-of-function’ mutation, occurring via a single-nucleotide change and ultimately resulting
in reduced G6PD production [10]. While G6PD Mahidol may confer a selective advantage to
parasitic infections, such as malaria, this genetic variation may also have other detrimental
affects to the immune response and may be implicated in the cell cycle of abnormal or
cancerous cells [9,10].
It has been found that the expression or enzyme activity of G6PD is elevated in multiple
tumors, including leukemia [11], gastrointestinal cancers [12], renal cell carcinomas [13],
colon cancers [14], breast cancers [15], endometrial carcinomas [16], prostate and liver
cancer [17,18]. Considering that ectopic G6PD expression increases the levels of
nicotinamide adenosine dinucleotide phosphate (NAPDH) and glutathione, G6PD may
promote the survival of tumor cells through maintenance of both extracellular pH and redox
potential [19,20], though little is known about how G6PD regulates cell proliferation and
apoptosis in tumors.
Persistent expression and excessive activation of STAT3 and STAT5 is apparent in
melanoma cells. Accordingly, STAT3 and STAT5 have become interesting potential target
molecules for the treatment of melanoma [21,22]. Our previous in vitro study demonstrated a
significant reduction in the P-STAT5/STAT5 ratio of A375-G6PD∆ cells following
knockdown of G6PD in A375 cells, while the P-STAT5/STAT5 ratio significantly increased
following overexpression of G6PD in the G6PD-knockdown A375 cells. This suggested that
G6PD promotes the proliferation of A375 cells and is associated with induction or activation
of STAT5. In addition, it has been found that STAT3 is persistently activated, and P-STAT3
expression is significantly elevated in A375 cells. STAT3 expression increased by five-fold
in G6PD-knockdown A375 cells compared to normal A375 cells, and P-STAT3 expression
levels in G6PD-knockdown A375 cells was 20% of that in A375-WT cells (unpublished
data). The activation of STAT3 is fast and transient under normal physiological conditions,
and it is strictly regulated [23]. These findings indicate that STAT3 and STAT5 play
important roles in mediating the biological characteristics of melanomas. However, the
underlying mechanism remains unclear.
The current study further explores the relationship and mechanism of action of G6PD and
melanoma cell proliferation and apoptosis using a mouse model of tumor formation. Human
dermal melanoma cells expressing the wild-type G6PD gene (A375-WT), G6PD-deficient
A375 cells (A375-G6PD∆), and A375-G6PD∆ cells with overexpression of normal G6PD
5. cDNA (A375-G6PD∆-G6PD-WT) and mutant G6PD cDNA (A375-G6PD∆-G6PD-G487A)
were administered to mice in order to compare the time of initial tumor formation, tumor
size, and pathological changes. In addition, the expression of G6PD and its activity, cell
cycle-related proteins, apoptosis-related proteins, and STAT3/STAT5 in tumor tissues were
determined in order to provide full documentation of the regulatory mechanisms involved
with in vivo melanoma growth associated with G6PD.
Methods
Cell culture
Human melanoma cell lines (A375) with knocked down G6PD genes (A375-G6PD∆) were
established from wild-type human dermal melanoma cell lines (A375-WT) (Cell Bank of the
Chinese Academy of Sciences) as previously described [24]. Wild-type and mutant-type
G6PD genes (G487A,G→A) were amplified using PCR and then cloned into a retroviral
vector (pBABEpuro) to yield the expression vectors pBABEpuro-G6PDWT and pBABE-
puro-G6PDG487A, respectively. The expression vectors were transfected into 293FT
package cells (R70007, Invitrogen, USA) using a retrovirus packaging kit (D6161, Takara,
Japan) to produce recombinant viruses. The recombinant retrovirus was used to infect the
A375-G6PD∆ cells and was subsequently screened for 7 days using puromycin (0.5 µg/mL)
(J593, Amreso, USA). Then, clones positive for puromycin-resistance were co-cultured in
G418 (200 µg/mL) and puromycin (0.25 µg/mL) to yield A375∆-WT and A375∆-G6PDA
cells exhibiting overexpression of G6PD. The passage and digestion of cells were similar to
those of the A375-G6PD∆ cells. Normal human epidermal melanocytes (HEM) were isolated
from primary-cultured melanocytes from the foreskin of healthy children [25] as a source for
mouse xenografts.
Establishment of a melanoma xenograft nude mouse model
A total of 25 4–5 w BALB/c strain nude mice (18–20 g) (Beijing HFK Bioscience Co, Ltd,
Beijing China) (Animal license number: SCXK 2009–0004; Beijing) were housed and raised
in the laboratory animal center of the Affiliated Cancer Hospital of Sun Yat-sen University.
The treatment and use of animals during the study was approved by the Animal Ethics
Committee of Sun Yat-sen University.
Mice were randomly assigned to 5 groups of 5 mice each: normal human epidermal
melanocytes (HEM), human dermal melanoma cells (A375-WT), G6PD-deficient A375 cells
(A375-G6PD∆), A375-G6PD∆ cells with overexpression of normal G6PD cDNA (A375-
G6PD∆-G6PD-WT), and A375-G6PD∆ cells with overexpression of mutant G6PD cDNA
(A375-G6PD∆-G6PD-G487A). Cells in the log-phase stage of growth were harvested and
digested into single cells using 0.25% pancreatin. Cells were washed with Dulbecco’s
Modified Eagle Medium (DMEM) without serum. A volume of 1 ml of each cell type
(1.5×106
/ml) was injected intradermally into the left axilla of the mice subjects. After
seeding, liquid absorption at the injection site, tumor growth (volume and weight), and mouse
survival were observed. Tumor volume was measured on days 5, 7, 12, 16, 19, 21, and 23
post-injection. On day 23, all mice were sacrificed, and tumors were isolated for
determination of weight and volume.
6. The largest (a) and smallest diameters (b) of each tumor were measured twice on days 5, 7,
12, 16, 19, 21, and 23 to estimate tumor volume (V) using the formula V = 0.52 × a2
× b [26].
Mean tumor volumes were used to plot tumor growth curves for each group of mice.
Immunohistochemistry
Immunohistochemistry was performed to determine associations between the G6PD
expression in various experimental groups, tumor growth, and pathological changes.
Tumor samples were fixed in 4% formaldehyde, embedded in paraffin wax, and then cut into
4 µm sections using a microtome. The sections were stained with hematoxylin and eosin
(HE).
Fixed tumor samples were prepared into 30 µm frozen sections and incubated with 2% goat
serum at 37°C for 20 min, followed by incubation with rabbit-anti-G6PD (1:500, Boster,
China) at 4°C overnight. After washing with PBS, the sections were incubated with
horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (HRP-IgG) at 37°C for 30 min
and colored with 3,3′-Diaminobenzidine (DAB) at room temperature. PBS was substituted
for the rabbit-anti-G6PD antibody in negative control subjects. The number of cells positive
for G6PD was then calculated.
Determination of G6PD activity
Tumor samples (20 mg) were homogenized with 5 ml of PBS at 4°C and centrifuged at 5000
r/min for 3 min. The supernatant (30 µl) was treated with a G6PD Activity Assay Kit reagent
(BioVision, USA) according to the manufacturer’s instructions. The optical density (OD
value, A1) of the positive control supplied by the kit was measured at 450 nm using a U-1800
ultraviolet spectrophotometer (Hitachi, Japan). After 30 min at 37°C the OD value (A2) was
measured a second time. The kit’s standard reagent, nicotinamide adenine dinucleotide
hydride (NADPH), was diluted 3 times, and the standard curve was plotted. A1 and A2 were
introduced into the standard curve to generate the total NADH (B). The G6PD activity was
then calculated using the formula: G6PD activity (mU/ml) = B/30 × V × dilution times.
Extraction of total RNA, reverse transcription, and quantitative real-time
PCR
Quantitative real-time PCR (qRT-PCR) was used to quantify the expression of G6PD mRNA
in the experimental groups. Tumor samples (60 mg) were ground under liquid nitrogen, lysed
with 1 ml of Trizol (Takara, Japan), and total RNA was extracted using Trizol (Invitrogen,
USA). Total RNA (2 µg) was added to the tumor extract with Moloney Murine Leukemia
Virus Reverse Transcriptase (MMLV-RT, Takara, Japan) to synthesize cDNA, and the
reverse transcript was used as the template for qRT-PCR using a Tower qRT-PCR system
(Analytic Jena, Germany). The qRT-PCR was conducted using 2×Mix SYBR Green I
(Biosea, USA) (10 µl), primer (0.25 µl, 10 pmol/L), template DNA (1 µl), and sterile water
(8.5 µl). All PCR reactions included initial denaturation and multiple cycles at (95°C for 3
min); 39 cycles at 95°C for 10 s, 55°C for 10 s, and 72°C for 30 s; followed by 95°C for 10 s,
65°C for 5 s, and a final 95°C for 15 s. The primer for each gene was synthesized by
Invitrogen (USA), and the sequences were determined (Table 1).
7. Table 1 Real-time PCR primer sequences
Gene Primer Sequence of primer Length of
sequence
(Base)
Length of
amplified
product (bp)
G6PD G6PD-F 5′TGAGCCAGATAGGCTGGAA3′ 19 225
G6PD-R 5′TAACGCAGGCGATGTTGTC3′ 19
CyclinD1CyclinD1-
F
5′CGGTAGTAGGACAGGAAGTT3′ 20 120
CyclinD1-
R
5′CTGTGCCACAGATGTGAAGT3′ 20
Fas Fas-F 5′GCCACCGACTTTAAGTTTGC3′ 20 102
Fas-R 5′CGAGCTCACTTCCTCATCCT3′ 20
STAT3 STAT3-F 5′CACCCGCAATGATTACAGTG3′ 20 126
STAT3-R 5′CGGTCTGACCTCTTAATTCG3′ 20
STAT5 STAT5-F 5′CACCCGCAATGATTACAGTG3′ 20 126
STAT5-R 5′CGGTCTGACCTCTTAATTCG3′ 20
β-actin β-actin-F 5′TGGCACCCAGCACAATGAA3′ 20 186
β-actin-R 5′CTAAGTCATAGTCCGCCTAGAAGCA3′ 25
Western blot
Tumor samples were lysed for 30 min in CytoBuster Protein Extraction Buffer (Novagen,
USA) and centrifuged at 12000 rpm. The supernatant was collected, total protein was
measured, and 50 µg was used for 10% sodium dodecyl sulfate polyacrylamide gel
electrophoresis (SDS-PAGE). The protein was then transferred to a nitrocellulose (NC)
membrane and sealed with Tris-Buffered Saline Tween-20 (TBST) containing 5% non-fat
milk powder. The membrane was subsequently incubated with rabbit anti-human STAT3, P-
STAT3, STAT5, and P-STAT5 proteins and mouse anti-human β-actin (1:500 to 1:1000,
Boster) at 4°C overnight. After washing in TBST, the membrane was incubated with HPR
conjugated secondary antibodies (1:6000) at 25°C, and the protein quantity was determined
using electrochemiluminescence (ECL) technique (BestBio, USA). The results were
photographed using the JS Gel Imaging System (Peiqing, China) and the grey density was
calculated using SensiAnsys software (Peiqing, China).
Statistical analysis
Statistical analyses were performed using SPSS statistical software version 13.0 (IBM, USA).
One-way analysis of variance (ANOVA) with 5 levels was used with a completely
randomized design, and the homogeneity of variance was tested. A q test (Student-Newman-
Keuls) was used to compare the differences between groups, and a rank sum test was
performed to randomly compare treatments. A P-value of less than 0.05 was considered
statistically significant (P < 0.05). Values were expressed as means ± standard deviation
(mean ± SD).
8. Results
Construction and identification of cell lines
After multiple freeze-thaw cycles (≥ 20), fluorescent protein [24] was stably expressed in
A375-G6PD∆, A375-G6PD∆-G6PD-WT, and A375-G6PD∆-G6PD-G487A cells, which
were morphologically normal (data not shown). The expression of G6PD mRNA, protein
quantity, and G6PD activity were all higher in A375 cells than those in normal HEM cells.
After G6PD knockdown, the expression of G6PD mRNA, G6PD protein, and enzyme
activity were significantly down-regulated (P < 0.01). In contrast, the G6PD mRNA, protein
expression, and enzyme activity were correspondingly elevated after the normal or G6PD
deficient gene was introduced into A375-G6PD∆ cells. A375-G6PD∆, A375-G6PD∆-G6PD-
WT, and A375-G6PD∆-G6PD-G487A cell lines were successfully established (Figure 1).
Figure 1 Identification of tumor cell lines. A, Determination of mRNA expression of G6PD
in various cell lines; B, Determination of expression of G6PD protein in various cell lines; C,
Determination of G6PD activity in various cell lines.
Effect of G6PD deficiency on tumor formation in nude mice injected with
A375 cell lines
Tumor formation was observed in 2 of 5 mice injected with A375-G6PD∆ cells. These
tumors were the last to form and grew the slowest among the treatment groups. The fastest
tumor growth was observed in the A375-WT cell group (Figure 2). Tumor formation was
observed 12 d after injection of cells with G6PD deficiency. Tumor sizes observed in the
G6PD deficiency group (A375-G6PD∆) were significantly smaller than those observed in the
other groups at 12 to 23 d post-injection (P < 0.05). Tumors presence was observed at 7 d
post-injection in A375-WT cells, and tumor size was significantly greater in this group than
in other groups at 16 days post-injection (P < 0.05).
Figure 2 Tumor formation and growth of 4 tumor cells in nude mice. After normal
human epidermal melanoma cells (HEM), human dermal melanoma cells (A375-WT),
G6PD-deficient A375 cells (A375-G6PD∆), A375-G6PD∆ cells with overexpression of
normal G6PD cDNA (A375-G6PD∆-G6PD-WT) and A375-G6PD∆ cells with
overexpression of mutant G6PD cDNA (A375-G6PD∆-G6PD-G487A) were injected into the
nude mice. The tumor size was measured on days 7, 12, 16, 19, 21 and 23 post-injection, and
the tumor volume growth curve (A) was plotted. The nude mice were photographed on day
23 (B), and then sacrificed. The tumors were isolated, and the weights (C) and volume of the
tumors were measured (D).
The smallest tumors at 23 d were observed in the G6PD deficiency group (P < 0.05), while
the largest tumors were in the A375-WT cell group (P < 0.05). There was no significant
difference in tumor weights between the A375-G6PD∆-G6PD-G487A and A375-G6PD∆-
G6PD-WT cell groups (P > 0.05), but both exhibited tumors that were larger than those
observed in the G6PD deficiency group (P < 0.05, Figure 2C).
Pathological staining results showed that tumors in the A375-WT cell group appeared to be
more malignant than those observed in other groups. Tumor cells taken from the A375-
G6PD∆ group exhibited the most benign changes compared with other groups (Additional
9. file 1: Table S1 and Additional file 2: Figure S1). No metastasis was observed in either the
liver or lungs in any groups, as determined via microscopic techniques.
G6PD expression and enzyme activity in nude mice tumor tissues
Immunohistochemistry demonstrated that the largest number of G6PD-positive cells were
present in the A375-WT cell group, while the amount of G6PD-positive cells was
significantly reduced in the A375-G6PD∆ cell group (Figure 3A, 3C). Similarly, qRT-PCR
indicated that the level of G6PD mRNA detected in the A375-WT cell group increased by
66-fold compared with that observed in the A375-G6PD∆ group (P < 0.05, Figure 3D). The
highest G6PD activity was observed in the A375-WT cell group, followed consecutively
lower activities observed in the A375-G6PD∆-G6PD-WT and A375-G6PD∆-G6PD-G487A
cell groups. The lowest G6PD activity was observed in the A375-G6PD∆ cell group (P <
0.01, Figure 3B), demonstrating the association of G6PD with melanoma formation and
growth in nude mice.
Figure 3 G6PD protein and mRNA expression and activity in tumor tissues of nude
mice from different experimental groups. A, Paraffin-embedded sections of the tumor
tissues of nude mice from different groups were made and the G6PD expression is
determined using immunohistochemistry (400×); B, Immunohistochemistry reveals the
G6PD-positive cells in tumor tissues; C, Changes of G6PD activity. n, a, b, c and d indicate
statistically significant differences, P < 0.05. n, vs. HEM cells group ; a, vs. A375-WT cells
group; b, vs. A375-G6PD∆ cells group; c, vs. A375-G6PD∆-G6PDWT cells group; d, vs.
A375- G6PD∆-G6PD-G487A cells group. D, mRNA expression of G6PD detected by qRT-
PCR.
Correlation of G6PD in tumor tissues with cell cycle protein expression
The qRT-PCR showed that mRNA levels of cyclin D1 increased by 31-fold in the A375-WT
cell group compared with the lowest observed levels in the A375-G6PD∆ cell group (P <
0.05, Figure 4D). Immunohistochemistry also suggested that the protein levels of Cyclin E,
p53, and S100A4 in the A375-G6PD∆ group decreased by 77%, 60%, and 74%, respectively,
compared with levels observed in the A375-WT group (Figure 4, Additional file 3: Figure S2,
Additional file 4 Figure S3, Additional file 5: Figure S4). Both the mRNA level of cyclin D1
and the protein levels of these three genes in A375-G6PD∆-G6PD-WT and A375-G6PD∆-
G6PD-G487A cell groups were determined to be partially recovered.
Figure 4 Determination of cell cycle-related genes in tumor tissues of nude mice from
different experimental groups. A, amounts of cyclin E-positive cells in tumor tissues. B,
amounts of p53-positive cells in tumor tissues. C, amounts of S100A4-positive cells in tumor
tissues. n, a, b, c and d indicate statistically significant differences (P < 0.05), n vs. HEM; a
vs. A375-WT; b vs. A375-G6PD∆; c vs. A375-G6PD∆-G6PDWT; d vs. A375-G6PD∆-
G6PD-G487A. D, mRNA expression of cyclinD1 detected by qRT-PCR.
Correlation of G6PD in tumor tissues with apoptosis-related protein
expression
Immunohistochemistry showed that the expression of apoptosis inhibitory factors Bcl-2 and
Bcl-xL was significantly down-regulated in the A375-G6PD∆ cell group, while the
10. expression of Fas protein increased by 72% compared with expression levels observed in the
A375-WT cell group (Figure 5, Additional file 6: Figure S5, Additional file 7: Figure S6,
Additional file 8: Figure S7). Protein expression data for Fas was also supported by the Fas
mRNA levels detected by qRT-PCR (Figure 5D). Again, results in A375-G6PD∆-G6PD-WT
and A375-G6PD∆-G6PD-G487A cell groups remained at moderate levels.
Figure 5 Determination of cell apoptosis genes in tumor tissues of nude mice from
different experimental groups. A, amounts of Bcl-2-positive cells in tumor tissues B,
amounts of Bcl-xl-positive cells in tumor tissues. C, amounts of Fas-positive cells in tumor
tissues and n, a, b, c and d indicate statistically significant differences (P < 0.05), n vs. HEM;
a vs. A375-WT; b vs. A375-G6PD∆; c vs. A375-G6PD∆-G6PDWT; d vs. A375-G6PD∆-
G6PD-G487A. D, mRNA expression of Fas detected by qRT-PCR.
Expression of STAT3 and STAT5 in tumor tissues
Western blot analysis results showed that STAT3, P-STAT3, STAT5, and P-STAT5 proteins
decreased by 47%, 65%, 38%, and 67%, respectively, in the A375-G6PD∆ group as
compared with results observed in the A375-WT group. Expressions of these proteins were
observed to be partially restored in the A375-G6PD∆-G6PD-WT and A375-G6PD∆-
G6PDG487A treated groups (Figure 6). The mRNA levels of STAT3 and STAT5 were
detected by qRT-PCR, providing further support for these results (Figure 6D, 6E). Data
presented in Figures 6B and 6C was generated from the average western blot data of all
animal tissues in each group.
Figure 6 Determination of STAT-3 and STAT-5 expression in tumor tissues. Expression
of STAT-3 and STAT-5 proteins and qualitative (A) and semi-quantitative analyses (B and
C) of their phosphorylations. 1, HEM; 2, A375-WT; 3, A375-G6PD∆; 4, A375-G6PD∆-
G6PDWT; 5, A375- G6PD∆-G6PD-G487A. STAT3 is persistently activated in A375-WT
cells. Compared with the HEM group, higher P-STAT3 expression is observed in the A375-
WT cells group. Following knockdown of G6PD in wild-type A375 cells, the expression of
both STAT3 and P-STAT3 decreases. The mRNA expression of STAT3 (D) and STAT5 (E).
Discussion
G6PD was previously shown to be highly expressed in human melanoma cells, exhibiting a
close relationship to the growth and proliferative phenotype of tumor cells [24], although no
in vivo study has confirmed this finding. Moreover, the underlying mechanism behind this
correlation may be important in developing a complete understanding of the pathogenesis of
melanoma and providing critical experimental evidence for the development of improved
clinical treatment options. In the current study, as expected, expression and activity of G6PD
showed a positive correlation with melanoma weight, growth, and differentiation. These
findings suggest that G6PD expression in the A375 cell line plays an important role in tumor
growth and proliferation. These findings are consistent with previous reports noting elevated
expression or activity of G6PD in tumor cells [11-18]. Furthermore, expression and activity
of G6PD was shown to be positively correlated with mRNA and protein expression of cyclins
D1 and E, p53, and S100A4. These findings suggest that G6PD can regulate cell cycle
through its effect on these factors, thus indirectly regulating melanoma growth.
11. Cyclin D1 accelerates the G1/S phase transition and promotes cell growth and division.
Cyclin D1 overexpression is considered to be an important factor in the promotion of tumor
occurrence and development, where it is also associated with reduced response to growth
factors [27,28]. Elevated cyclin D1 gene expression in melanoma cells revealed that cyclin
D1 was the key protein promoter of A375 cell proliferation and its overexpression can cause
cell proliferation out of control which promotes tumor malignancy. Cyclin E is synthesized
initially in the G1 phase, and expression increases in mid-G1 phase. Thereafter, cyclin E
expression sharply increases and reaches its peak at the G1/S junction, followed by rapid
disappearance in the S phase [29]. Immunohistochemical staining revealed few cyclin E-
positive cells in the HEM group and significantly elevated cyclin E expression in the A375-
WT group, suggesting that cyclin E is an important factor in melanoma occurrence and
consistent with report by Bales E [30]. Furthermore, expression levels of cyclin D1 and E are
positively correlated with expression of G6PD and its activity. These findings have indicated
the vital regulation role of G6PD in uncontrolled cell proliferation and resultant malignant
transformation in melanomas.
The tumor suppressor protein p53, a crucial regulator of the cell cycle in multicellular
organisms, is also not expressed in certain cancers, including skin melanomas. It is, however,
overexpressed in invasive melanomas with a high proliferative index [31,32]. Elevated p53
expression observed in A375 cells correlated well with high G6PD activity, suggesting that
protein p53 is promoted by G6PD, which is consistent with study by Murtas et al. [33]. Jiang
P found that the protein p53 affect the growth and invasion of melanoma cells in nude mice
by interfering with the pentose-phosphate pathway in tumor cells [34]. These results suggest
p53 may affect glucose metabolism in human melanoma when the activity of G6PD is
interfered. S100A4 is a protein closely related to cellular differentiation and tumor
occurrence, metastasis, and prognosis. S100A4 binds to the p53 protein and restricts the
function of the G-S restriction point in the cell cycle. It may also be associated with
uncontrolled entry in the M phase by bypassing the G2-M restriction point [35,36]. The
present study showed that the level of S100A4-positive cells correlated well with p53, G6PD
activity, and tumor growth, suggesting that G6PD affects p53 activity, thus also impacting
melanoma occurrence and metastasis by regulation of S100A4 expression.
G6PD also showed a positive correlation to mRNA and protein expression of cell apoptosis
inhibitory factors Bcl-2 and Bcl-xl and a negative correlation to Fas. Fas can be used as a
marker to escape apoptosis by malignant cells. The immune escape mechanism employed by
malignant melanomas involves actively attacking cytotoxic T lymphocytes (CTL) that
express Fas to escape from apoptosis [37]. The Bcl-2 protein family affects apoptosis by
regulating mitochondrial and endoplasmic reticulum stress-induced apoptotic pathways
[38,39]. The proportion of apoptosis-inhibitory protein (Bcl-2) and apoptosis-promoting
protein (Bax) determines the rate of apoptosis occurrence if cells are stimulated by apoptotic
signals. Higher Bax/Bcl-2 ratios result in relatively higher sensitivity to CD95/Fas-mediated
apoptosis [40]. In the present study, lower Fas expression in the A375-WT group and
elevated Fas expression in the A375-G6PD∆ group with the G6PD knockout corresponded to
low expression levels of apoptosis-inhibitory proteins Bcl-2 and Bcl-xL in the A375-G6PD∆
group. Similarly, high expression was observed in the A375-WT group, suggesting that
G6PD may regulate cell apoptosis of melanoma through apoptosis-related factors Fas, Bcl-2,
and Bcl-xL. Cumulatively, G6PD has a varied mechanism of action, affecting the levels of
numerous other proteins associated with cell cycle regulation, thus promoting the
development of malignancies.
12. The STAT pathway is well-down for its association with proliferation of malignant
melanoma, escape from apoptotic signals, tumor invasion and angiogenesis [41]. More
importantly, activation of STAT is the key factor for development of melanoma [42];
silencing of STAT3 using short hairpin RNA (shRNA) inhibits the growth of melanoma in
tumor-bearing mice [43].
STAT3 can down-regulate Bcl-xL expression and induce apoptosis in human melanoma
A2058 and Jw cell lines [44]. In metastatic melanoma, the expression of phosphorylated
STAT3 is positively correlated with Bcl-xL expression [45]. Suppression of STAT5 protein in
human melanoma A375 cells significantly down-regulates Bcl-2 expression [46], while
activation of STAT5 up-regulates Bcl-xL expression [42]. This indicates that STAT5 is an
important anti-apoptotic factor [22,46]. Similarly, the present study indicated that STAT3 and
STAT5 were positively correlated with Fas expression and negatively correlated with
expression of Bcl-2 and Bcl-xL. Leslie et al. [47] demonstrated that Cyclin D1 mRNA levels
were significantly increased in tumor cell lines that had excessive activation of the STAT3
signaling pathway, while mutagenesis of STAT3 binding sites within the cyclin D1 promoter
region significantly inhibited the transcription of the Cyclin D1 gene. Cyclin D1 level was
also positively correlated with STAT3 level in our study. Altogether, it suggests that the
STAT3/STAT5 pathway could possibly be involved in the mechanism behind cell cycle
abnormalities and cell apoptosis regulation.Further mechanistic studies, however, will be
required to verify this mechanism.
Conclusion
G6PD is involved in the growth, proliferation, and apoptosis of neoplastic tumors in nude
mice models of melanoma. Furthermore, the tumor malignancy was observed to be in accord
with G6PD protein expression. Higher expression of G6PD protein promoted the survival and
proliferation of neoplastic tumors in nude mice models of melanoma (A375 cells) through the
up-regulation of cyclin D1 and cyclin E, P53 and S100A4 protein expression. In this model, a
deficiency of G6PD proteins in A375-G6PD∆ cells promoted cell apoptosis through down-
regulation of the expressions of Bcl-2 and Bcl-xL and up-regulations of the expression of
Fas. The correlation of G6PD expression and tumor growth corresponded with
STAT3/STAT5 expression, although further experiments will be required to confirm the
direct regulatory role of G6PD on this pathway. Because of the broad affects of G6PD, it may
provide an excellent target for the development of improved treatment methods and
prognostic indicators for melanoma patients.
Competing interest
The authors declare that they have no competing interest.
Authors’ contributions
TH and ZZ carried out the immunoassays. CZ and QT carried out real-time PCR and Western
blot assay. BL setup the nude mice model. LC and TCcarried out the G6PD activity assay.
YS and YZ carried out the design of the study and drafted the manuscript. All authors read
and approved the final manuscript.
13. Funding sources
This work was supported by grants from the National Natural Science Foundation of China
(No. 30860322; No. 81160246), the Talented Person Foundation of Yunnan Province (No.
2007PY01-13).
References
1. Cappellini MD, Fiorelli G: Glucose-6-phosphate dehydrogenase deficiency. Lancet
2008, 371(9606):64–74.
2. Beutler E: G6PD deficiency. Blood 1994, 84(11):3613–3636.
3. WHO Working Group: Glucose-6-phosphate dehydrogenase deficiency. Bull World
Health Organ 1989, 67(6):601–611. 67.
4. Minucci A, Moradkhani K, Hwang MJ, Zuppi C, Giardina B, Capoluongo E: Glucose-6-
phosphate dehydrogenase (G6PD) mutations database: review of the “old” and update
of the new mutations. Blood Cells Mol Dis 2012, 48(3):154–165.
5. Zhao X, Li Z, Zhang X: G6PD-MutDB: a mutation and phenotype database of glucose-
6-phosphate (G6PD) deficiency. J Bioinforma Comput Biol 2010, 8(Suppl 1):101–109.
6. Yang Y, Zhu Y, Li D, Li Z, Lu H, Wu J, Tang J, Tong S: Characterization of glucose-6-
phosphate dehydrogenase deficiency and identification of a novel haplotype
487G>A/IVS5-612(G>C) in the Achang population of Southwestern China. Science in
China Series C, Life sciences / Chinese Academy of Sciences 2007, 50(4):479–485.
7. Louicharoen C, Patin E, Paul R, Nuchprayoon I, Witoonpanich B, Peerapittayamongkol C,
Casademont I, Sura T, Laird NM, Singhasivanon P: Positively selected G6PD-Mahidol
mutation reduces Plasmodium vivax density in Southeast Asians. Science 2009,
326(5959):1546–1549.
8. Panich V: Glucose-6-phosphate dehydrogenase deficiency. Part 2. Tropical Asia. Clin
Haematol 1981, 10(3):800–814.
9. Hedrick PW: Population genetics of malaria resistance in humans. Heredity 2011,
107(4):283–304.
10. Casanova JL, Abel L, Quintana-Murci L: Human TLRs and IL-1Rs in host defense:
natural insights from evolutionary, epidemiological, and clinical genetics. Annu Rev
Immunol 2011, 29:447–491.
11. Batetta B, Pulisci D, Bonatesta RR, Sanna F, Piras S, Mulas MF, Spano O, Putzolu M,
Broccia G, Dessi S: G6PD activity and gene expression in leukemic cells from G6PD-
deficient subjects. Cancer Lett 1999, 140(1–2):53–58.
14. 12. Wang J, Yuan W, Chen Z, Wu S, Chen J, Ge J, Hou F: Overexpression of G6PD is
associated with poor clinical outcome in gastric cancer. Tumour biology: the journal of the
International Society for Oncodevelopmental Biology and Medicine 2012, 33(1):95–101.
13. Langbein S, Frederiks WM, zur Hausen A, Popa J, Lehmann J, Weiss C, Alken P, Coy
JF: Metastasis is promoted by a bioenergetic switch: new targets for progressive renal
cell cancer. Int J Cancer 2008, 122(11):2422–2428.
14. Van Driel BE, Valet GK, Lyon H, Hansen U, Song JY, Van Noorden CJ: Prognostic
estimation of survival of colorectal cancer patients with the quantitative histochemical
assay of G6PDH activity and the multiparameter classification program CLASSIF1.
Cytometry 1999, 38(4):176–183.
15. Polat MF, Taysi S, Gul M, Cikman O, Yilmaz I, Bakan E, Erdogan F:
Oxidant/antioxidant status in blood of patients with malignant breast tumour and
benign breast disease. Cell Biochem Funct 2002, 20(4):327–331.
16. Philipson KA, Elder MG, White JO: The effects of medroxyprogesterone acetate on
enzyme activities in human endometrial carcinoma. J Steroid Biochem 1985,
23(6A):1059–1064.
17. Nna E, Tothill IE, Ludeman L, Bailey T: Endogenous control genes in prostate cells:
evaluation of gene expression using ‘real-time’ quantitative polymerase chain reaction.
Medical principles and practice: international journal of the Kuwait University, Health
Science Centre 2010, 19(6):433–439.
18. Rao KN, Elm MS, Kelly RH, Chandar N, Brady EP, Rao B, Shinozuka H, Eagon PK:
Hepatic hyperplasia and cancer in rats: metabolic alterations associated with cell
growth. Gastroenterology 1997, 113(1):238–248.
19. Kobayashi M, Fujita I, Itagaki S, Hirano T, Iseki K: Transport mechanism for L-lactic
acid in human myocytes using human prototypic embryonal rhabdomyosarcoma cell
line (RD cells). Biol Pharm Bull 2005, 28(7):1197–1201.
20. Maly K, Hochleitner B, Uberall F, Loferer H, Oberhuber H, Doppler W, Grunicke H:
Mechanism and biological significance of the Ha-ras-induced activation of the
Na+/H(+)-antiporter. Adv Enzym Regul 1990, 30:63–74.
21. Messina JL, Yu H, Riker AI, Munster PN, Jove RL, Daud AI: Activated stat-3 in
melanoma. Cancer control: journal of the Moffitt Cancer Center 2008, 15(3):196–201.
22. Mirmohammadsadegh A, Hassan M, Bardenheuer W, Marini A, Gustrau A, Nambiar S,
Tannapfel A, Bojar H, Ruzicka T, Hengge UR: STAT5 phosphorylation in malignant
melanoma is important for survival and is mediated through SRC and JAK1 kinases. J
Investig Dermatol 2006, 126(10):2272–2280.
23. Bromberg J: Stat proteins and oncogenesis. J Clin Investig 2002, 109(9):1139–1142.
15. 24. Li D, Zhu Y, Tang Q, Lu H, Li H, Yang Y, Li Z, Tong S: A new G6PD knockdown
tumor-cell line with reduced proliferation and increased susceptibility to oxidative
stress. Cancer Biother Radiopharm 2009, 24(1):81–90.
25. Eisinger M, Marko O: Selective proliferation of normal human melanocytes in vitro
in the presence of phorbol ester and cholera toxin. Proc Natl Acad Sci U S A 1982,
79(6):2018–2022.
26. Niu G, Heller R, Catlett-Falcone R, Coppola D, Jaroszeski M, Dalton W, Jove R, Yu H:
Gene therapy with dominant-negative Stat3 suppresses growth of the murine melanoma
B16 tumor in vivo. Cancer Res 1999, 59(20):5059–5063.
27. Raisova M, Bektas M, Wieder T, Daniel P, Eberle J, Orfanos CE, Geilen CC: Resistance
to CD95/Fas-induced and ceramide-mediated apoptosis of human melanoma cells is
caused by a defective mitochondrial cytochrome c release. FEBS Lett 2000, 473(1):27–32.
28. Sauter ER, Yeo UC, von Stemm A, Zhu W, Litwin S, Tichansky DS, Pistritto G, Nesbit
M, Pinkel D, Herlyn M, et al: Cyclin D1 is a candidate oncogene in cutaneous melanoma.
Cancer Res 2002, 62(11):3200–3206.
29. Arooz T, Yam CH, Siu WY, Lau A, Li KK, Poon RY: On the concentrations of cyclins
and cyclin-dependent kinases in extracts of cultured human cells. Biochemistry 2000,
39(31):9494–9501.
30. Bales E, Mills L, Milam N, McGahren-Murray M, Bandyopadhyay D, Chen D, Reed JA,
Timchenko N, van den Oord JJ, Bar-Eli M, et al: The low molecular weight cyclin E
isoforms augment angiogenesis and metastasis of human melanoma cells in vivo. Cancer
Res 2005, 65(3):692–697.
31. Bergman R, Shemer A, Levy R, Friedman-Birnbaum R, Trau H, Lichtig C:
Immunohistochemical study of p53 protein expression in Spitz nevus as compared with
other melanocytic lesions. Am J Dermatopathol 1995, 17(6):547–550.
32. Miracco C, Santopietro R, Biagioli M, Lazzi S, Nyongo A, Vatti R, Luzi P: Different
patterns of cell proliferation and death and oncogene expression in cutaneous malignant
melanoma. J Cutan Pathol 1998, 25(5):244–251.
33. Murtas D, Piras F, Minerba L, Ugalde J, Floris C, Maxia C, Demurtas P, Perra MT, Sirigu
P: Nuclear 8-hydroxy-2'-deoxyguanosine as survival biomarker in patients with
cutaneous melanoma. Oncol Rep 2010, 23(2):329–335.
34. Jiang P, Du W, Wang X, Mancuso A, Gao X, Wu M, Yang X: p53 regulates
biosynthesis through direct inactivation of glucose-6-phosphate dehydrogenase. Nat Cell
Biol 2011, 13(3):310–316.
35. Cajone F, Sherbet GV: Stathmin is involved in S100A4-mediated regulation of cell
cycle progression. Clin Exp Metastasis 1999, 17(10):865–871.
36. Sherbet GV, Lakshmi MS: S100A4 (MTS1) calcium binding protein in cancer growth,
invasion and metastasis. Anticancer Res 1998, 18(4A):2415–2421.
16. 37. Bullani RR, Wehrli P, Viard-Leveugle I, Rimoldi D, Cerottini JC, Saurat JH, Tschopp J,
French LE: Frequent downregulation of Fas (CD95) expression and function in
melanoma. Melanoma Res 2002, 12(3):263–270.
38. Brunelle JK, Letai A: Control of mitochondrial apoptosis by the Bcl-2 family. J Cell
Sci 2009, 122(Pt 4):437–441.
39. Heath-Engel HM, Chang NC, Shore GC: The endoplasmic reticulum in apoptosis and
autophagy: role of the BCL-2 protein family. Oncogene 2008, 27(50):6419–6433.
40. Raisova M, Hossini AM, Eberle J, Riebeling C, Wieder T, Sturm I, Daniel PT, Orfanos
CE, Geilen CC: The Bax/Bcl-2 ratio determines the susceptibility of human melanoma
cells to CD95/Fas-mediated apoptosis. J Investig Dermatol 2001, 117(2):333–340.
41. Barton BE, Karras JG, Murphy TF, Barton A, Huang HF: Signal transducer and
activator of transcription 3 (STAT3) activation in prostate cancer: Direct STAT3
inhibition induces apoptosis in prostate cancer lines. Mol Cancer Ther 2004, 3(1):11–20.
42. Hatiboglu MA, Kong LY, Wei J, Wang Y, McEnery KA, Fuller GN, Qiao W, Davies
MA, Priebe W, Heimberger AB: The tumor microenvironment expression of p-STAT3
influences the efficacy of cyclophosphamide with WP1066 in murine melanoma models.
International journal of cancer Journal international du cancer 2012, 131(1):8–17.
43. Manuel ER, Blache CA, Paquette R, Kaltcheva TI, Ishizaki H, Ellenhorn JD, Hensel M,
Metelitsa L, Diamond DJ: Enhancement of cancer vaccine therapy by systemic delivery
of a tumor-targeting Salmonella-based STAT3 shRNA suppresses the growth of
established melanoma tumors. Cancer Res 2011, 71(12):4183–4191.
44. Farley J, Smith LM, Darcy KM, Sobel E, O’Connor D, Henderson B, Morrison LE,
Birrer MJ: Cyclin E expression is a significant predictor of survival in advanced,
suboptimally debulked ovarian epithelial cancers: a Gynecologic Oncology Group
study. Cancer Res 2003, 63(6):1235–1241.
45. Krasilnikov M, Ivanov VN, Dong J, Ronai Z: ERK and PI3K negatively regulate
STAT-transcriptional activities in human melanoma cells: implications towards
sensitization to apoptosis. Oncogene 2003, 22(26):4092–4101.
46. Zhuang L, Lee CS, Scolyer RA, McCarthy SW, Zhang XD, Thompson JF, Hersey P:
Mcl-1, Bcl-XL and Stat3 expression are associated with progression of melanoma
whereas Bcl-2, AP-2 and MITF levels decrease during progression of melanoma.
Modern pathology: an official journal of the United States and Canadian Academy of
Pathology, Inc 2007, 20(4):416–426.
47. Wellbrock C, Weisser C, Hassel JC, Fischer P, Becker J, Vetter CS, Behrmann I,
Kortylewski M, Heinrich PC, Schartl M: STAT5 contributes to interferon resistance of
melanoma cells. Curr Biol 2005, 15(18):1629–1639.
17. Additional files
Additional_file_1 as DOC
Additional file 1: Table S1 Pathological observations of neoplasm tumor in nude bearing
melanoma model after injection of five types of cells (showed by HE staining).
Additional_file_2 as TIFF
Additional file 2: Figure S1 HE staining of tumor tissues produced by injection of 5 types of
cells.
Additional_file_3 as TIFF
Additional file 3: Figure S2 Immunohistochemical staining of cyclin E protein in tumors
formed by injection of 4 types of cells.
Additional_file_4 as TIFF
Additional file 4: Figure S3 Immunohistochemical staining of p53 protein in tumors
produced by injection of 4 types of cells.
Additional_file_5 as TIFF
Additional file 5: Figure S4 Immunohistochemical staining of S100A4 protein in tumors
produced by injection of 4 types of cells.
Additional_file_6 as TIFF
Additional file 6: Figure S5 Immunohistochemical staining of Fas protein in tumors
produced by injection of 4 types of cells.
Additional_file_7 as TIFF
Additional file 7: Figure S6 Immunohistochemical staining of Bcl-2 protein in tumors
produced by injection of 4 types of cells.
Additional_file_8 as TIFF
Additional file 8: Figure S7 Immunohistochemical staining of Bcl-xL protein in tumors
produced by injection of 4 types of cells.