Targeting Inhibit The Expression Of C-FLIP By MiR-150/miR-504/miR-519d Regulates And Controls Proliferation、Apoptosis And Autophagy In Colorectal Carcinoma Cells

Part 1.To analyze and detect the expression of c-FLIP in colorectal caicinoma tissue.Objective: To analyze and detect the expression of c-FLIP in colorectal caicinoma tissue.Methods: lay experiment research for tissue chip of colorectal caicinoma,immunohistochemical and fluorescence in situ bybridization methods were used.Results:The significant expression of c-FLIP was found in colorectal caicinoma tissue.Conclusion:The obvious expression of c-FLIP in colorectal caicinoma tissue predicts c-FLIP can play a promotion for colorectal caicinoma cells,we can modulate this action through target-suppressing c-FLIP.Part 2.To investigate the influence of proliferation、apoptosis and autophagy in colorectal carcinoma by target-regulating the expression of c-FLIP with miR-150/miR-504/miR-519 d.Objective:To investigate the influence of proliferation、apoptosis and autophagy in colorectal carcinoma by target-regulating the expression of c-FLIP with miR-150/miR-504/miR-519 d.Methods:First obstain nine miRNAs genes which can target-suppress c-FLIP through predicting micro RNA targets on website and then test or verify on further,analyze the relationship between nine miRNAs and c-FLIP in SW48 cells by real-time PCR..we acquired three miRNAs which were negative correlation with c-FLIP.Luciferase analyses was used for testing the hypothesis that c-FLIPm RNA might be a target of three miRNAs.Finally,obversing the function of tumor cells with over-expression of miRNA.Results:C-FLIP was negative correlation with has-miR-519d、has-miR-504 and has-miR-150,and was further proved through the down-modulated of c-FLIP when tumor cells were transfected with has-miR-519 d 、 has-miR-504 and has-miR-150,and at the same time,luciferase analyses confirmed they all can target-suppress c-FLIP respectively,over-expressed miRNA could decrease cell viability、inhibit cell proliferation and migrating、accelerate cell apoptosis and autophagy.The influence of cell function by over-expressed 3 connecting miRNA was small.Conclusions: C-FLIP was negative correlation with has-miR-519d、has-miR-504、has-miR-150(p<0.05),over-expression of has-miR-519d、has-miR-504 and has-miR-150 can target-suppress c-FLIP respectively(p<0.05)、decrease cell viability(p<0.05)、inhibit cell proliferation and migrating(p<0.05)、accelerate cell apoptosis and autophagy(p<0.05).The influence of cell function by over-expressed 3 connecting miRNA was small,perhaps there was antagonism factor in space construction.