Expermental Studies On Interactions Between MicroRNAs And CDH1mRNA In The EMT Process

The leading cause of death in patients with cancer are due to metastasis of tumor cells caused by complications. Tumor cells metastasis prOocess involving mesenchymal cells turn into epithelial cells,which called epithelial-mesenchymal transition process (EMT).Gene CDH1,which encoding E-cadherin protein,plays a key role in the EMT, and has a high levels expressiont in the epithelial cells.When E-cadherin downregulation or missing will reduce cell adhesion strength and result in increasing cells activity,at the same times, make tumor cells through the basement membrane invating the surrounding tissue.Investigating the microRNAs (miRNAs) interaction with the target gene CDH1can help people to understand the process of the tumor diseases happening,and provide some therapeutic direction for clinical medicine. This paper mainly aims at cervical cancer cell, research the interaction mechanism between the target gene CDH1and miRNAs, mainly complete following three aspects work:(1) First we use predictive softwares Targetscan, miRanda, Pictar Search the potential miRNAs which have binding site at the3’UTR of CDH1.Finally,we confirm mir-9, mir-23b, mir-92a, mir-340miRNAs as Research Targets. Because literature experimental data has indicate mir-9can downregulate the E-cadherin protein,it is regard as a reference in our experiment.(2) Using the method of molecular biology experiment,first we construct mir-9, mir-23b, mir-92a and CDH1plasmid and transfect them into HeLa cells (human tumor cells) and293T cells (human normal cells,this paper as the control group).Secondly, through the dual luciferase report system,we get some quantitative measurement results which indicate some selected miRNAs indeed downregute gene CDH1.Through wound healing experiments we observe the tumor cell activity from the cellular level.Our data indicate that mir-9,mir-23b,mir-92a indeed inhibit the CDH1expression, thereby increasing the migration of cancer cells.(3) By comparison with the existing regulatory mechanisms of miRNAs and CDH1, combined with the experimental results of this paper, we propose a potential signal path in the process of EMT in cancer cells that regulated by mir-9,mir-23b.