微RNA-186在人肝细胞肝癌中的表达及作用
发布时间：2018-10-15 18:27
【摘要】：目的:检测微RNA-186(miRNA-186,miR-186)在肝细胞肝癌(hepatocellular carcinoma,HCC)组织和细胞中的表达水平,以及其对肝癌SMMC-772细胞生物学特性的影响。方法:采用实时荧光定量PCR法检测34对HCC组织及其对应癌旁组织中miR-186的表达情况,同时检测4株HCC细胞(Hep G2、Hep3B、SMMC-7721和BEL-7402)以及正常肝细胞株HL-7702中miR-186的表达情况。将携带有miR-186-模拟物(mimics)和miR-186-抑制物(inhibitor)的真核表达载体转染至SMMC-7721细胞后,分别采用CCK-8法、FCM法以及Transwell小室迁移和侵袭实验检测细胞增殖、凋亡、迁移和侵袭能力的变化。分别采用实时荧光定量PCR和蛋白质印迹法进一步检测miR-186对靶基因ROCK1的作用。结果:miR-186在HCC组织中的表达量为0.034±0.033,明显低于其在癌旁组织中的表达量0.077±0.056(P0.001)。临床病理学特征分析结果显示,HCC组织中miR-186的表达量与肿瘤大小和TNM分期有关(P值均0.05)。miR-186在HCC细胞中的表达量均明显低于正常肝细胞(P值均0.05),其中以在SMMC-7721细胞中的表达水平最低。与对照组相比,miR-186-mimics组细胞在各时间点的增殖活性均明显降低,而miR-186-inhibitor组细胞在各时间点的增殖活性均明显升高(P值均0.05);miR-186-mimics组细胞的凋亡率明显升高,而miR-186-inhibitor组细胞凋亡率明显降低(P值均0.05);miR-186-mimics组细胞的迁移和侵袭能力明显降低,而miR-186-inhibitor组细胞的迁移和侵袭能力明显增强(P值均0.05)。过表达miR-186能够降低ROCK1 m RNA及蛋白的表达水平,抑制miR-186表达则会上调ROCK1 m RNA及蛋白的表达水平(P值均0.05)。结论:miR-186在HCC组织和细胞中均低表达,并能够通过下调ROCK1的表达来抑制HCC细胞的增殖、迁移和侵袭,同时促进细胞凋亡。
[Abstract]:Aim: to investigate the expression of microRNA-186 (miRNA-186,miR-186) in hepatocellular carcinoma (hepatocellular carcinoma,HCC) and its effect on the biological characteristics of SMMC-772 cells. Methods: the expression of miR-186 in 34 pairs of HCC and its adjacent tissues was detected by real-time fluorescence quantitative PCR. The expression of miR-186 in four HCC cells (Hep G2Hep3BnSMMC-7721 and BEL-7402) and the normal hepatocyte line HL-7702 was also detected. After transfection of eukaryotic expression vector carrying (mimics) and miR-186- inhibitor (inhibitor) into SMMC-7721 cells, the changes of cell proliferation, apoptosis, migration and invasion were detected by CCK-8 assay, FCM assay and Transwell chamber migration and invasion assay, respectively. The effect of miR-186 on target gene ROCK1 was further detected by real-time fluorescence quantitative PCR and Western blotting. Results: the expression of miR-186 in HCC was 0.034 卤0.033, which was significantly lower than that in paracancerous tissues (0.077 卤0.056). The results of clinicopathological analysis showed that the expression of miR-186 in HCC was related to tumor size and TNM stage (P < 0. 05). The expression of miR-186 in HCC cells was significantly lower than that in normal hepatocytes (P = 0. 05), especially in SMMC-7721 cells. Compared with the control group, the proliferative activity of miR-186-mimics group was significantly lower than that of control group, while the proliferative activity of miR-186-inhibitor group was significantly increased at each time point (P < 0. 05), and the apoptosis rate of miR-186-mimics group was significantly higher than that of control group. The apoptosis rate of miR-186-inhibitor group was significantly lower than that of miR-186-mimics group (P < 0. 05), but the migration and invasion ability of miR-186-mimics group was significantly lower than that of miR-186-inhibitor group (P < 0. 05). Overexpression of miR-186 could decrease the expression of ROCK1 m RNA and protein, but inhibit the expression of miR-186 could up-regulate the expression of ROCK1 m RNA and protein (P < 0 05). Conclusion: the expression of miR-186 is low in HCC tissues and cells, and it can inhibit the proliferation, migration and invasion of HCC cells by down-regulating the expression of ROCK1, and promote the apoptosis of HCC cells.
【作者单位】： 重庆医科大学附属第一医院肝胆外科;
【分类号】：R735.7
[Abstract]:Aim: to investigate the expression of microRNA-186 (miRNA-186,miR-186) in hepatocellular carcinoma (hepatocellular carcinoma,HCC) and its effect on the biological characteristics of SMMC-772 cells. Methods: the expression of miR-186 in 34 pairs of HCC and its adjacent tissues was detected by real-time fluorescence quantitative PCR. The expression of miR-186 in four HCC cells (Hep G2Hep3BnSMMC-7721 and BEL-7402) and the normal hepatocyte line HL-7702 was also detected. After transfection of eukaryotic expression vector carrying (mimics) and miR-186- inhibitor (inhibitor) into SMMC-7721 cells, the changes of cell proliferation, apoptosis, migration and invasion were detected by CCK-8 assay, FCM assay and Transwell chamber migration and invasion assay, respectively. The effect of miR-186 on target gene ROCK1 was further detected by real-time fluorescence quantitative PCR and Western blotting. Results: the expression of miR-186 in HCC was 0.034 卤0.033, which was significantly lower than that in paracancerous tissues (0.077 卤0.056). The results of clinicopathological analysis showed that the expression of miR-186 in HCC was related to tumor size and TNM stage (P < 0. 05). The expression of miR-186 in HCC cells was significantly lower than that in normal hepatocytes (P = 0. 05), especially in SMMC-7721 cells. Compared with the control group, the proliferative activity of miR-186-mimics group was significantly lower than that of control group, while the proliferative activity of miR-186-inhibitor group was significantly increased at each time point (P < 0. 05), and the apoptosis rate of miR-186-mimics group was significantly higher than that of control group. The apoptosis rate of miR-186-inhibitor group was significantly lower than that of miR-186-mimics group (P < 0. 05), but the migration and invasion ability of miR-186-mimics group was significantly lower than that of miR-186-inhibitor group (P < 0. 05). Overexpression of miR-186 could decrease the expression of ROCK1 m RNA and protein, but inhibit the expression of miR-186 could up-regulate the expression of ROCK1 m RNA and protein (P < 0 05). Conclusion: the expression of miR-186 is low in HCC tissues and cells, and it can inhibit the proliferation, migration and invasion of HCC cells by down-regulating the expression of ROCK1, and promote the apoptosis of HCC cells.
【作者单位】： 重庆医科大学附属第一医院肝胆外科;
【分类号】：R735.7
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