Mir-181a Negatively Regulates Kras To Inhibit The Proliferation And Migration Of Cholangiocarcinoma Cell Line Qbc939

Objective: To investigate whether miR-181a can regulate the expression of Kras to affect the proliferation and migration of cholangiocarcinoma cell line QBC939. Methods: QBC939 cells were divided into D group (cells without treatment), Dz group (cells transfected with negative control), Dm group (cells transfected with miR-181a inhibitor), and Dy group Dy (cells transfected with miR-181a mimics). The morphology of cells was observed by Hoechst staining, and the expression of Kras mRNA and miR-181a was determined by RT-PCR. The expression of Kras protein was detected by Western blot, and the invasion ability of QBC939 cells was detected by transwell assay. Clone formation assay and MTT assay were used to detect cell cloning ability and viability, and the dual luciferase reporter gene assay was performed. Results: Cells in Dz group and D group were arranged in a regular manner, while cells in Dm group were tightly arranged with increased quantity. Besides, cells in Dy group were sparsely arranged and fractured with increased loss of nuclear membrane. The expression of Kras mRNA was the lowest in Dy group and the highest in Dm group, while the expression of miR-181a was the opposite. The transfection of miR-181a could reduce Kras activity (P<0.05), but the effect on the mutated gene was not significant (P>0.05). Western blot showed that Kras protein expression in Dm group was higher than that in other groups (P<0.05), and the expression in Dz and D group was higher than that in Dy group (P<0.05). Transwell assay showed that the overall invasion ability of cells in Dm group was enhanced compared with other three groups (all P<0.05). Compared with other groups, the invasion ability of cancer cells was the weakest and the number of cells under microscope was the lowest in Dy group (all P<0.05). The clone formation assay and MTT assay showed that the number of monoclonal populations and cell viability in Dy group were the lowest. The monoclonal formation rate of QBC939 cells in Dm group, Dz group and D group showed an uptrend, which was significantly higher than that in Dy group (all P<0.05). Conclusion: Overexpression of miR-181a could inhibit proliferation and migration of cholangiocarcinoma cell line QBC939, which may be mediated by targeting and inhibiting the expression of Kras.