miR-148a对皮肤鳞状细胞癌迁移能力的影响

Effect of miR-148a on the migrationability of skin squamous cell carcinoma

目的探讨miR-148a对皮肤鳞状细胞癌(CSCC)迁移能力的影响。方法收集2017年1~6月于新乡医学院第一附属医院确诊的50例CSCC患者手术切除癌组织和对应癌旁正常组织,采用实时定量聚合酶链反应检测CSCC患者癌组织和癌旁正常组织中miR-148a和转化生长影响因子2(TGIF2)mRNA表达。取CSCC细胞株A431培养24 h后随机分为阴性对照组、miR-148a mimics组和miR-148a inhibitor组,分别转染阴性对照、miR-148a mimics和miR-148a inhibitor质粒,转染后培养48 h,采用Western blot法检测各组A431细胞中TGIF2蛋白表达;分别于转染后0、36、72 h进行划痕愈合实验检测各组A431细胞迁移能力。结果CSCC组织中miR-148a相对表达量显著低于癌旁正常组织(P<0.05),CSCC组织中TGIF2 mRNA相对表达量显著高于癌旁正常组织(P<0.05)。miR-148a在CSCC组织中的表达与患者的性别和临床分期无关(P>0.05),与患者的年龄、病理分化和淋巴结转移有关(P<0.05)。miR-148a mimics组A431细胞中TGIF2蛋白表达量显著低于阴性对照组(P<0.05),miR-148a inhibitor组A431细胞中TGIF2蛋白表达量显著高于miR-148a mimics组和阴性对照组(P<0.05)。转染后36、72 h,miR-148a mimics组A431细胞划痕愈合率低于阴性对照组(P<0.05),miR-148a inhibitor组A431细胞划痕愈合率高于miR-148a mimics组和阴性对照组(P<0.05)。结论CSCC组织中miR-148a表达显著下调,TGIF2表达显著上调,CSCC A431细胞中miR-148a过表达可使TGIF2蛋白表达下降,从而抑制细胞迁移。

Objective To investigate the influence of miR-148a on the migration ability of skin squamous cell carcinoma(CSCC).Methods The cancer tissues and corresponding adjacent normal tissues of 50 CSCC patients who were diagnosed in the First Affiliated Hospital of Xinxiang Medical University from January to June 2017 were collected,and the expression of miR-148a and transforming growth interacting factor 2(TGIF2)mRNA in cancer tissues and adjacent normal tissues were detected by real-time quantitative polymerase chain reaction.The CSCC A431 cells were cultured for 24 h,then the cells were randomly divided into negative control group,miR-148a mimics group and miR-148a inhibitor group.The negative control,miR-148a mimics and miR-148a inhibitor plasmids were transfected into A431 cells of above three groups respectively.The expression of TGIF2 protein in A431 cells in the three groups was detected by Western blot at 48 hours after transfection;the migration ability of A431 cells in the three groups was detected by scratch healing experiment at 0,36,72 h after transfection.Results The relative expression of miR-148a in CSCC tissues was significantly lower than that in the adjacent normal tissues(P<0.05),and the relative expression of TGIF2 mRNA in CSCC tissues was significantly higher than that in the adjacent normal tissues(P<0.05).The expression of miR-148a in CSCC tissues was not related to the gender and clinical stage of patients(P>0.05),and it was related to the age,pathological differentiation and lymph node metastasis(P<0.05).The expression of TGIF2 protein in A431 cells in the miR-148a mimics group was significantly lower than that in the negative control group(P<0.05);the expression of TGIF2 protein in A431 cells in the miR-148a inhibitor group was significantly higher than that in the miR-148a mimics group and negative control group(P<0.05).At 36,72 hours after transfection,the scratch healing rate of A431 cells in the miR-148a mimics group was lower than that in the negative control group(P<0.05);the scratch healing rate of A431 cells in the miR-148a inhibitor group was higher than that in the miR-148a mimics group and negative control group(P<0.05).Conclusion The expression of miR-148a is down-regulated in CSCC tissues,and the expression of TGIF2 is up-regulated in CSCC tissues.The overexpression of miR-148a in CSCC A431 cells can inhibit the cell migration behavior of cells by decreasing the expression of TGIF2 protein.