The Confirmation Of 3’UTR Region Function Mutation Loci Of Sporadic Colorectal Cancer Candidate Genes

This study through searching colorectal cancer related information in the published literatures and Ensembl database, And using the common target gene prediction software to predict3 ’UTR region specific miRNA targets, of 12 candidate genes,the results showed TGFBR2 (Rs17026336) gene and SMAD3 gene (Rs746978) combined with good specificity of micrornas are respectively hsa miR 3191-3 p and has miR 3200-5 p, so determine SMAD3, TGFBR2 as candidate genes of colorectal cancer. Through constructed the 3 ’UTR polymorphism loci of candidate genes of wild type and mutant expression vector, adopted the method of dual luciferase report gene analysis of candidate genes with the target relationship of micrornas, the results show that the hsa-miR-3191-3 p has significantly correlation with TGFBR2 gene expression level, and hsa-miR-3200-5 p was not significant effect on SMAD3 gene.In this study, compared hsa-miR-3191-3 p in normal colon cell line (NCM460) and different stages of tumor cell lines (sw480, colo320, lovo, HT29) expression level by real time quantitative PCR.Results show that hsa-miR-3191-3 p expression is significantly higher in normal colonic cell lines than the department of cancer cells (sw480, colo320, lovo, HT29).Conclusion:3 ’UTR region of TGFBR2 Rs17026336 may with hsa-miR-3191-3 p interaction affect the development of colorectal cancer, the results in order to further determine whether TGFBR2 gene as candidate genes of colorectal cancer provides experimental basisCAPNI, as the important proteinase of cytoplasm, is a kind of soluble calcium depende nt neutral protease. Its plays an influential role on cytoskeleton remodeling, signal transductio n and cell death. Studies have shown that there was an relationship between CAPNI gene and cancer, which found that CAPNI had a middle or lower expression in the renal clear cell carc inoma. In this study, we detected CAPNI mutated to LoF(Loss-of-Function)among a case of colorectal cancer tissue samples. In recent years, there is no in-depth study about the relations hip between colorectal and CAPN1.In recent years, there is no in-depth study about the relatio nship between colorectal and CAPN1.In this study, we constructed pEGFP C2 and pIRES2-EGFP expression vectors of CAP Nl gene, analyzed the expression of CAPN1 between normal colon cell lines and different sta ges of tumor cell lines such as sw480, HT-29, colo320 and lovo by Western Blot, and results shown that the normal colon NCM460 cell lines had a lower expression compared with tumor cell lines, Which providing a theory basis for furthering determine the CAPNI gene is the ca ndidate genes of colorectal cancer.