Effects Of Suppressing The Expression Of MiRNA-21 On The Proliferation, Apoptosis And Invasion Abilities In Human Cholangiocarcinoma Cells And A Study Of Its Target Gene

Cholangiocarcinomas (CCAs) are aggressive cancers, with high mortality and poor survival rates. Only radical surgery offers patients some hope of cure. However, most patients are not surgical candidates because of late diagnosis secondary to relatively poor accuracy of diagnostic means. Rencent yeas, researcher discovered that microRNAs (miRNAs) were involved in every cancer examined and played an very important role. miRNA-21 was overexpressed in CCA tissue and was regarded as an oncogene. this study inhibited the expression of miRNA-21 in human cholangiocarcinoma cell line RBE cells, examined the changes of biological behavior in RBE cells and identified potential regulating target mRNA.Objective To discuss the effects of suppressing the expression of miRNA-21 in human cholangiocarcinoma cell line RBE cells on the proliferation and apoptosis of RBE cells and its potential regulating target mRNA.Method The human cholangiocacinoma cell line,RBE cells was divided into 4 groups: natural growth cells, lipo group(treated with Lipofectin), negative control group(transfected with negative counterpart), miRNA-21 inhibited group(transfected with miRNA-21 inhibitors). The proliferation change was detected by MTT method and the apoptosis was detected by flow cytometer(FCM). The invasion ability of RBE cells was analysed by vitro invasion assay(transwell). The changes in the expression of RECK and maspin mRNA and protein were detected by real-time RT-PCR and Western blotting respectively.Results After transfecting miRNA-21 inhibitors, the expression of miRNA-21in RBE cells was decreased to 0.0014 fold compared with the original cell line. MTT results showed that the proliferation was inhbited obviously(P<0.05)and FCM results showed that apoptosis of 4 groups cells were2.29%(Cell group), 2.43%(lipo group), 2.89%(miRNA-21-NC group), 19.35%(miRNA-21-IN group). The apoptosis significantly accelerated when the expression of miRNA-21 was suppressed. The cell counting of Transwell inserts under the microscope were 54.0±2.1(cell), 45.4±3.2(lipo), 40.8±2.4(miRNA-21-NC), 19.2±1.5(miRNA-21-IN). the invasion ability was inhbited significantly when the miRNA-21 was kocked down. The mRNA and protein of RECK and maspin were over-expressed after transfecting the miRNA-21 inhibitors in RBE cells (P<0.05).Conclusion The proliferation and invasion ability was inhibited and the apoptosis was accelerated in RBE cells by suppressing the expression of the miRNA-21.RECK and maspin was the target gene of miRNA-21 by participating in the process of regulation in RBE cells.