MicroRNA-7对HCCC9810细胞增殖、迁移和侵袭的研究

目的探讨MicroRNA-7(miR-7)对于肝内胆管癌细胞HCCC9810增殖、迁移、侵袭能力的影响。方法利用实时定量PCR对于肝内胆管癌病人的癌组织和癌旁组织,以及正常肝内胆管上皮细胞和肝内胆管癌细胞HCCC9810中miR-7表达水平的验证;培养人肝内胆管癌细胞HCCC9810,将其分为研究组(miR-7 mimics)和阴性对照组(miR-7 NC),分别进行miR-模拟物以及miR-阴性序列的转染,效率验证后,通过CCK-8实验,EDU实验,克隆形成实验,以及划痕和Transwell实验分别验证两组细胞的增殖、迁移和侵袭的能力。结果肝内胆管癌病人中癌组织miR-7的表达水平低于癌旁组织,肿瘤细胞HCCC9810 miR-7表达水平低于正常胆管上皮细胞;细胞转染效率验证显示,miR-7 mimics组HCCC9810细胞中miR-7的相对表达水平远高于miR-7 NC组,差异具有统计学意义(P<0.05);与miR-7NC相比,miR-7 mimics组细胞的增殖、迁移以及侵袭能力被明显抑制,差异均有统计学意义(均P<0.05)。结论miR-7在肝内胆管癌中表达水平下调,而且过表达miR-7能够抑制肿瘤细胞的增殖、迁移以及侵袭过程。

Study on the effects of MicroRNA-7 in proliferation,migration and invasion of HCCC9810 cells

Objective To investigate the effect of MicroRNA-7(miR-7)on the proliferation,migration and invasion of intrahepatic cholangiocarcinoma cells HCCC9810.Methods Real-time quantitative PCR was used to verify the expression level of miR-7 in the cancer tissue and adjacent normal tissue of patients with cholangiocarcinoma,as well as in normal biliary epithelial cells and intrahepatic cholangiocarcinoma cells HCCC9810.Human cholangiocarcinoma cells HCCC9810 were cultured and divided into experimental group(miR-7 mimics)and negative control group(miR-7 NC).Transfection of miR-mimics and miR-negative sequences was performed,respectively.After efficiency verification,the proliferation,migration and invasion of the two groups of cells were verified by CCK-8 assay,EDU assay,colony formation assay,and scratch and transwell assay,respectively.Results The expression level of miR-7 in cancer tissues of patients with intrahepatic cholangiocarcinoma is lower than adjacent tissues,and the expression level of miR-7 in intrahepatic cholangiocarcinoma cells is lower than normal bile duct epithelial cells.The verification of cell transfection efficiency showed that the relative expression level of miR-7 in HCCC9810 cells in miR-7 mimics group was significantly higher than that in miR-7 NC group,and the difference was statistically significant(P<0.05).Compared with the miR-7 mimics group,cell proliferation,migration,and invasion ability were significantly inhibited,and the differences were statistically significant(all P<0.05).Conclusion The expression level of miR-7 is down-regulated in intrahepatic cholangiocarcinoma,and overexpression of miR-7 can inhibit the proliferation,migration and invasion of tumor cells.