Clinical Development of Biomarker To Detect Oral Carcinoma In Relation To Genetic Polymorphism At MMP-9

Premalignant/potentially malignant oral lesion and condition such as oral submucous fibrosis are known to be transformed into oral cancer. The malignant transformation is often associated with genetic polymorphism which is reflected by the altered expression of proteins related to ECM degradation, fibroblast proliferation and angiogenesis. Protein such as metallo matrixproteinase-9 that loose substrate binding activity due to SNP at Q279R polymorphism at active site results in the degradation of type-4 collagen in basement membrane. The prime objective of this study is to develop biomarker for early detection of oral carcinoma, in relation to MMP-9 polymorphism at Q279R. As a method, Genomic DNA were extracted from blood samples (5ml, from OSF and control subjects) for genotypic analysis using restriction fragment length polymorphism (RFLP) subsequently after amplification through polymerase chain reaction (PCR). The expressions of MMP-9 Polymorphism in transcriptional and post-translational levels were analyzed by RT-PCR and Western blot respectively. Statistical validation was performed by estimating P-values using student-t test and odds ratio analysis. It was found that the odd ratio of R/R compared to Q/Q was 1.75 (0.41 -7.48) and p-value was 0.575. The RT-PCR analysis of MMP-9 has shown the significant up-regulation of transcription compared to that of normal p-value 0.024, observed in student t-test. Post- translational analysis using Western Blot has shown relatively high intensity of active MMP-9 in comparison to the normal (p-value 0.001) by using student t-test. The study is concluded on the basis of significant up regulation of active MMP-9 (Q279R-polymorfized; 82kDa) in patients than that of healthy control, we can conclude by establishing this polymerized protein as a biomarker for the early detection of oral submucous fibrosis to prevent the oral carcinoma