The Research On TCF7L2Influenced The Mechanism Of Insulin Secretion

Aims:To research TCF7L2influenced the mechanism of insulinsecretion. To explore the interaction of transcription factor TCF7L2andGPR40gene promoter. To study the effects of lentivirus mediated TCF7L2RNAi on expression of GPR40in β TC6cells.Methods：(1) We use chromatin immunoprecipitation (ChIP) inβTC6cells. Specific TCF7L2antibody was involved in, theimmunoprecipitated DNA fragments were amplified through PCR using thespecific GPR40primers.(2)TCF7L2RNAi lentivirus vector wasconstructed and was used to transfect βTC6cells. RNAi efficiency wasassessed by Real-time PCR and Western blotting.(3) Determine theglucose stimulate insulin secretion (GSIS) on control group and interferegroup.(4)The expression of GPR40was also assessed by Real-time PCRand Western blotting in TCF7L2stable interfere，s βTC6cells.Results：(1)GPR40gene specific fragment was detected in theDNA fragments immunoprecipitated by TCF7L2antibody.(2)TCF7L2RNAi lentivirus vector was successfully constructed. Lentivirus mediatedTCF7L2RNAi down-regulated TCF7L2mRNA and protein expression in βTC6cells， Real-time PCR showed that TCF7L2mRNA was decreased72%and78%compared to control group（P<0.05）.(3)The interfere group,GSIS was decreased compared to control group（P<0.05）.(4) GPR40mRNAand protein expression were descend in TCF7L2stable interfere，sβ TC6cells， Real-time PCR showed that GPR40mRNA decreased78%and88%compared to control group（P<0.05）.Conclusion： The results confirm that TCF7L2was binded toGPR40promoter inβTC6cells and transcription factor TCF7L2mayinvolved in transcriptional regulation of GPR40gene. Lentivirus mediatedTCF7L2RNAi can result in long time silencing TCF7L2gene, downregulate the expression of GPR40in β TC6cells. TCF7L2influencedinsulin secretion by GPR40.