沉默EMI2基因抑制肝内胆管癌HuCCT1细胞的增殖和上皮间质转化

Silencing EMI2 gene inhibits proliferation and epithelied-mesenchymal transformation of intrahepatic cholangiocarcinoma HuCCT1 cells

目的:探索早期有丝分裂抑制子2(early mitotic inhibitor 2,EMI2)在肝内胆管癌(intrahepatic cholangiocarcinoma,iCCA)中的表达及生物学功能,并探讨EMI2对iCCA细胞HuCCT1增殖、侵袭及上皮-间质转化(epithelial-mesenchymal transformation,EMT)的影响。方法:生物信息学技术检测EMI2在胆管癌中的表达情况,构建沉默EMI2的HuCCT1稳转株细胞(shEMI2),观察其对HuCCT1细胞增殖、侵袭和EMT的影响。结果:生物信息学技术显示EMI2在胆管癌组织中高表达,Western Blot和RT-PCR检测EMI2敲低效率。CCK-8验证shEMI2组的OD_(450)值要低于shCtrol组(P<0.05),且shEMI2组细胞克隆形成数目(26±2)低于shCtrol组(57±4)。侵袭结果显示,shEMI2组侵袭细胞数(39±4.92)较shCtrol组(195±14.00)少。细胞周期实验显示,shEMI2组较shCtrol组G_(2)/M期的细胞比例减少,处于G_(1)期的细胞比例增多(P<0.05)。细胞凋亡实验表明,shEMI2组的细胞凋亡率提高(P<0.05)。Western Blot和免疫荧光结果提示,沉默EMI2可以促进E-cadherin蛋白上调,N-cadherin和Vitmentin蛋白含量下调。结论:沉默EMI2基因抑制iCCA细胞增殖、侵袭、迁移和EMT的过程。

Objective:To explore the expression and biological function of early mitotic inhibitor 2(EMI2)in intrahepatic cholangiocarcinoma(iCCA)tissues.To investigate the effects of EMI2 on the proliferation,invasion and epithelial-mesenchymal transformation of iCCA cells HuCCT1.Methods:Bioinformatics technique was used to detect the expression of EMI2 in bile duct carcinoma.Gene silencing technique was used to silence EMI2,and its effects on proliferation,invasion,migration and EMT of HuCCT1 cells were observed.Results:Bioinformatics showed that EMI2 was highly expressed in cholangiocarcinoma tissues,while Western Blot and RT-PCR were ineffective in detecting EMI2.CCK-8 showed that the OD_(450) value of shEMI2 group was lower than that of shCtrol group(P<0.05),and the number of shEMI2 cell clone formation(26±2)was lower than that of shCtrol group(57±4).Invasive results showed that the number of invasion in shEMI2 group was(39±4.92)and in shCtrol group was(195±14.00).Cell cycle test showed that compared with shCtrol group,shEMI2 group of G_(2)/M cells decreased and G_(1) cells increased(P<0.05).The apoptosis experiment showed that the apoptosis rate of shEMI2 group was increased(P<0.05).Western Blot and immunofluorescence results suggested that silencing EMI2 could promote the up-regulation of E-cadherin protein and down-regulation of N-cadherin and Vitmentin protein.Conclusion:Silencing EMI2 gene inhibites the proliferation,invasion,migration and epithelied-mesenchymal transformation of intrahepatic cholangiocarcinoma.