【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。更多还原

【Abstract】 Objective To investigate the effect and mechanism of miR-203 a-3 p 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-203 a-3 p group(transfected with miR-203 a-3 p 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-203 a-3 p and zinc finger transcription factor(SLUG).Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-203 a-3 p 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-203 a-3 p,SLUG mRNA,SLUG protein and epithelial mesenchymal transition related protein between the two groups were not significantly different,(P >0.05).In miR-203 a-3 p group,the invasion and migration ability was significantly decreased.The expression levels of SLUG mRNA,SLUG protein,N-cadherin and Vimentin in miR-203 a-3 p group were significantly decreased,while the expression levels of miR-203 a-3 p,E-cadherin and Occludin were significantly increased(P<0.05).Conclusion MiR-203 a-3 p may inhibit the migration and invasion of HCC cells by inhibiting EMT via regulating SLUG.更多还原