Information related to cancer, staging, biomarkers, genomics, transcriptomics and further research gaps are enlisted

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2. Detection of cancer by using salivary
biomarkers
Presented By; Menail Sajid

3. Contents
• Cancer and its staging
• Saliva in cancer detection
• History and success rate
• Characteristics and composition of saliva
• Saliva as diagnostic fluid
• Methods of collection of saliva
• Technologies for discovering biomarkers
• Genomic, transcriptomic and proteomic technologies
• Why saliva?
• Objectives of study
• Further research areas
• Conclusion and References
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4. Cancer
• Abnormal growth that can invade nearby tissues.
• Second most deadly disease.
• Most common type of cancer found in Pakistan is breast cancer.
• Occur due to multiple reasons like;
• Oncogene
• BCL-2
• Caspase
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5. Staging of Cancer
Stage What does it mean?
Stage 0 No cancer, chances to get
tumorous
Stage I Small cancer, confined to one
area (benign)
Stage II & III Cancer invade tissues or
lymph nodes by growing
large in size
Stage IV Metastasis, Cancer spreads to
different organs
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6. Saliva in detection
• A multi-constituent oral fluid.
• To use saliva for detection purpose, prerequisites are;
• Discovering Biomarker
• Check sensitivity and specificity of Biomarker
• Saliva can be used for detection of diseases like;
• HIV infection, Hepatitis A, B & C, Cancer, Cardiovascular
diseases, metabolic diseases
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7. History and success rate
• Salivary biomarkers for cancer detection has been used from last two
decades.
• There is 90% survival rate in case of patients detected with salivary
biomarkers.
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8. Characteristics of Saliva
• Clear, acidic(pH=6.0-7-0).
• Secreted from parotid, submandibular and sublingual
glands.
• Secreted 1-1.5 L per day.
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9. Composition of saliva
• Saliva consists of;
• water, electrolytes, organic compounds, inorganic ions,
protein/polypeptides and enzymes like amylase
• Different types of protein present in saliva and act as
biomarker are;
• albumin, statherin, cystatins, lactoferrin, proline rich
proteins, histatins, transferrin, and α-amylase
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10. Saliva as diagnostic fluid
Sampling
Centrifugation (2000rpm for 05minute, 14000rpm
for 20minutes)
Add RNase/proteinase inhibitor
Discard pellet and keep supernatant
2D gel electrophoresis/mass spectrometry
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11. Continue
• Biomarkers(cancer) obtained from saliva are;
• M2BP, Profilin, IL-1B, IL-8, MRP14, and catalase
• Having 90% sensitivity and 80% specificity for cancer diagnosis.
• Bioinformatics tool used is, PubMed
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12. Methods of collection
• Saliva can be collected by two ways;
• Unstimulated
• Draining, spitting, suction or swab
• Stimulated
• saliva is collected by providing the patients
with a stimulant agent, such as, citric acid,
paraffin, or a gum base.
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13. Technologies for discovering biomarkers
• Salivary biomarker by genomic technology
• Salivary biomarker by transcriptomic technology
• Salivary biomarker by proteomic technology
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14. Genomic Changes/Genome Technology
• Totality of genetic material.
• Change in cellular DNA occur in case of cancer like;
• Point mutation
• Deletion
• Amplifications
• Methylations
• HPV presence
• Genome extracted from saliva shows;
• P53 gene mutation
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15. Continue
• Mitochondrial DNA mutation.
• Hyper methylation of promoter region of certain genes.
• Amplification of D1 gene.
• Presence of HPV and Epstein Barr viruses genome
sequences.
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16. Transcriptomic/Altered mRNA transcript
• Related to mRNA expression level.
• Seven mRNA molecules found up-regulated in cancerous patients.
• These seven mRNA molecules are;
• IL8(Immune response, replication, cell cycle arrest)
• IL1B(Proliferation, inflammation, apoptosis)
• DUSPI(protein modification)
• H3F3A(DNA binding activity)
• OAZ1(Polyamine biosynthesis)
• S100P(Protein and calcium ion binding)
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17. Proteomics/Alter protein markers
• Large scale study of proteins.
• Protein level elevate in case of cancer.
• 46 proteins are found at significantly different levels like;
• Higher amount of Definsin.
• Elevated concentration of interleukin 8.
• Proteomic of saliva is studied by using Luminex xMAP
technology.
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18. Limitations/Why saliva??
1. Lack of definitive biomarker for specific disease.
2. Lack of easy and inexpensive sampling method with
minimal discomfort.
3. Lack of an accurate, easy-to-use, and portable platform
to facilitate early detection.
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19. Continue
• Saliva contain large amount of compounds.
• Easily accessible.
• Patient comfort.
• Safe to handle.
• Low chances of transmission than blood.
• Easy to store.
• Saliva does not clot.
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20. Objectives of this study
To compare and Propose:
• Easy sample collection for diagnostic purpose.
• Safe and economic extraction of body fluid.
• Early detection of cancer.
• Involvement of even non-technical person for sample collection.
• Replacement of blood/urine tests for diagnostic applications.
• Less or no requirement of any special instrument for sample
collection.
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21. Conclusion
• Salivary biomarkers represent a promising, non-invasive approach for
cancer detection and specifically the oral cancer. It is area of strong
research interest. Along with all these, it can be an excellent
replaceable of blood and urine tests for detection purposes not only for
cancer but also for other lethal disorders.
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22. Further Research Areas
• Lack of standardization of saliva sample collection, processing, and
storage.
• Difference between changing concentration of salivary biomarkers in
case of cancer and other oral inflammatory diseases.
• Biological variance and physiological changes affecting oral cancer
salivary biomarkers.
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23. References
• Am J Dent. 2009 Aug; 22(4): 241–248. Saliva: An emerging biofluid
for earlu detection of diseases.
• Cheng et al. Clinical and Translational Medicine 2014, 3:3
• Curr Oral Health Rep (2014) 1:133–141
• Rosas et al
• 2010; 4: 172–178.
• Published online 2010 Aug 27. doi: 10.2174/1874210601004010172
• PMCID: PMC3111739
• PMID: 21673842
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24. Continue
• Salivary Markers for Oral Cancer Detection
• Anastasios K. Markopoulos,*,1 Evangelia Z.
Michailidou,1 and Georgios Tzimagiorgis2
• T. Radhika et al. / Journal of Oral Biology and Craniofacial Research 6
(2016) S51–S54S52
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