The Effect Of MicroRNA-133 On The Migration And Invasion Of Cholangiocarcinoma Cells

Objective:With the in-depth study of MicroRNA in recent years,it has been found that it is widely involved in cell proliferation,apoptosis and differentiation,and plays an important role in the pathological and physiological changes of cells.Cholangiocarcinoma is a highly aggressive biliary tract tumor originating from bile duct epithelial cells,and its morbidity and mortality are increasing worldwide.This study aimed to investigate the expression of MicroRNA-133(miR-133)and its different matures in cholangiocarcinoma QBC939 cells line.The specific miRNA interference reagent(Anti-mi R-133)was used to block the expression of endogenous miR-133 in cholangiocarcinoma QBC939 cells line,and then the role of miR-133 gene expression in the migration and invasion of cholangiocarcinoma cells was studied,and preliminary discussion of possible regulatory mechanisms.Methods:The cholangiocarcinoma cells QBC939 was seeded in DMEM containing 20% fetal bovine serum and cultured.Real-time quantitative PCR(RT-PCR)technology was used for total RNA extraction,reverse transcription to obtain c DNA,and used to analyze the expression of miR-133 and its different matures(miR-133 b,miR-133a-3p,miR-133a-5p)in cholangiocarcinoma cells QBC939.Single factor analysis of variation was used to compare the expression of different matures.Based on the results of RT-PCR,the mature with the highest expression level was selected for later interference experiments.Select specific miRNA interference reagents(Anti-miR-133)to block the expression of endogenous mi R-133 in cholangiocarcinoma cells QBC939,and QBC939 cells were chosen and divided into three groups: natural growth(blank control group),cell groups transfection with negative sequence(negative control group)and cell groups transfection with anti-miR-133(interference group).The expression of miR-133 and c-Met in the QBC939 cells of each group were measured by RT-PCR,and Single factor analysis of variation was used to compare whether the expression differences of miR-133 and c-Met genes in the three groups were statistically significant.Finally,the effects of miR-133 on the migration and invasion of QBC939 cells was analyzed by Transwell assay.Results:RT-PCR was used to detect the expression of mi R-133 different mature miR-133b(0.74 ± 0.07),miR-133a-3p(0.99 ± 0.12),miR-133a-5p(1.00 ± 0.06)in cholangiocarcinoma cells QBC939,Among them,miR-133a-5p had the highest expression in cholangiocarcinoma cells QBC939(P <0.05).RT-PCR experiments confirmed that the expression of miR-133a-5p in the interference group QBC939 cells was(0.70±0.08),the expression of miR-133a-5p in the negative control group QBC939 cells was(1.43±0.34),which was significantly lower in the interference group than the control group(P <0.05);Similarly,the expression of c-Met in the interference group QBC939 cells was(0.09 ± 0.02),the expression of c-Met in the blank control group QBC939 cells was(1.01 ± 0.07),the interference group was significantly lower than the control group(P <0.01).Transwell experiments confirmed that the migration(69.5±8.3)and invasion(19.3±3.3)abilities of QBC939 cells in the interference group were significantly lower than that of the negative and blank control groups(both P<0.01).conclusion:The low expression of MicroRNA-133 in vitro can significantly reduce the expression of c-Met gene,and can inhibit the migration and invasion ability of cholangiocarcinoma cells QBC939.mi R-133 may have a potential oncogene role in cholangiocarcinoma by regulating the tumor-associated gene c-Met,thereby affecting tumor cell proliferation,migration and invasion,miR-133 may become a new target for the regulation of cholangiocarcinoma oncogene expression.