miR-203a-3p通过SLUG调控上皮间质化抑制肝癌细胞的迁移及侵袭

摘要/Abstract

摘要： 目的探究miR-203a-3p对肝癌细胞迁移和侵袭的影响及其作用机制。方法体外培养HepG2细胞,将实验分为对照组(人肝癌细胞)、miRNA-NC组(转染阴性对照序列)和miR-203a-3p组(转染miR-203a-3p mimic)。Transwell和细胞划痕实验用以检测细胞侵袭和迁移能力;双荧光素酶实验用以验证miR-203a-3p和锌指转录因子(SLUG)的靶向关系;实时荧光定量聚合酶链式反应(qRT-PCR)用以检测miR-203a-3p和SLUG mRNA表达;蛋白质免疫印迹(WB)用以检测SLUG蛋白以及上皮间质转化相关蛋白E型钙黏蛋白(E-cadherin)、闭合蛋白(Occludin)、N型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)的表达;裸鼠成瘤实验用以检测转染后细胞体内成瘤能力。结果与对照组相比,miRNA-NC组细胞侵袭和迁移能力、miR-203a-3p的表达、SLUG mRNA和蛋白的表达、上皮间质转化相关蛋白的表达均差异无统计学意义(P>0.05);miR-203a-3p组细胞侵袭和迁移能力、SLUG mRNA和蛋白的表达、N-cadherin、Vimentin蛋白表达显著降低,miR-203a-3p的表达、E-cadherin、Occludin蛋白表达显著升高(P<0.05)。结论 miR-203a-3p可能靶向调控SLUG,抑制上皮间质转化过程,从而抑制肝癌细胞迁移及侵袭。

关键词: miR-203a-3p, 锌指转录因子, 肝癌细胞, 上皮间质转化, 迁移, 侵袭

Abstract: Objective To investigate the effect and mechanism of miR-203a-3p on the migration and invasion of hepatocellular carcinoma (HCC) cells. Methods HepG2 cells were cultured in vitro and divided into control group (human HCC cell line), miRNA-NC group (transfected with mimics control) and miR-203a-3p group (transfected with miR-203a-3p mimic). Transwell and scratch test were used to assess the invasion and migration ability. Luciferase reporter assay was used to verify the targeting relationship between miR-203a-3p and zinc finger transcription factor (SLUG). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-203a-3p and SLUG mRNA. Western blot (WB) was used to detect the expression levels of SLUG protein, E-cadherin, Occludin, N-cadherin and Vimentin. The tumorigenicity ability of transfected cells was detected in nude mice. Results There was no significant difference in invasion and migration ability of cells between control group and miRNA-NC group, and the expression levels of miR-203a-3p, SLUG mRNA, SLUG protein and epithelial mesenchymal transition related protein between the two groups were not significantly different, (P>0.05). In miR-203a-3p group, the invasion and migration ability was significantly decreased. The expression levels of SLUG mRNA, SLUG protein, N-cadherin and Vimentin in miR-203a-3p group were significantly decreased, while the expression levels of miR-203a-3p, E-cadherin and Occludin were significantly increased (P<0.05). Conclusion MiR-203a-3p may inhibit the migration and invasion of HCC cells by inhibiting EMT via regulating SLUG.

Key words: MiR-203a-3p, Zinc finger transcription factor, Hepatocellular carcinoma cells, Epithelial mesenchymal transition, Migration, Invasion

张国柄, 徐江海, 王东风. miR-203a-3p通过SLUG调控上皮间质化抑制肝癌细胞的迁移及侵袭[J]. 肝脏, 2021, 26(12): 1339-1343.

ZHANG Guo-bing, XU Jiang-hai, WANG Dong-feng. miR-203a-3p inhibits migration and invasion of HCC cells by regulating epithelial mesenchymal transition via SLUG[J]. Chinese Hepatolgy, 2021, 26(12): 1339-1343.

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