The Experimental Study On Target CDH1Regulation By MiR-106a、miR-376a And MiR-485-5p In Hela Cells

Epithelial to mesenchymal transition (EMT) is the process of epithelial cells with polarity transformed to mesenchymal cells with athletic ability. Tumor metastasis and invasion are closely related with EMT process,a complex process under regulation of a variaty of biomolecules (such as proteins,nucleic acids,ect). A large number of experiments have shown that:microRNA (miRNA) paly a very important role in the process of tumor EMT, as one of the hot topic in the current study of tumor invasion and migration mechanism.This paper mainly focus on EMT process of Hela cells, Firstly, using bioinformatic method, we theoretically screened out related miRNAs that paly an important role in Hela cells EMT:miR-106a, miR-376a and miR-485-5p. Then using molecular biological method, we study the regulatory mechanism of predicted miRNAs on target gene CDH1. The main work and achievements are as follow:(1) Firstly, using bioinformatic method (miRanda、TargetScan、FindTar、PicTar and miRBase), we theoretically predicted a number of miRNAs that could base pair with target gene CDH1.We screened out miR-106a、miR-376a and miR-485-5p which have negative regulation effect on CDH1, inhibit the expression of cadherin, have impact on Hela cells invasion and metastasis, as the main objects of research.(2) Secondly, applying molecular biological experiments, we constucted four plasmids containing miR-106a, miR-376a, miR-485-5p, and CDH13’UTR region respectively; then cotransfected both miRNA and mRNA plasmids into Hela cells, by flurescence detector measure the fluorescence expression levels of CDH13’UTR plasmid.(3) Finally, analysing the molecular experiment data:after transfection of miRNA plasmid, the fluorescence level of CDH1plasmid, the E-cadherin expression level are all down regulated significantly, the invasion ability of Hela cells are enhanced. The experiment result is consistent with theoretical prediction, prove that miR-106a, miR-376a and miR-485-5p can target regulate CDH13’UTR, enhance cells metastatic and invasive ability.