上皮细胞转化序列2诱导肝癌细胞上皮-间充质转化促进侵袭转移

Epithelial cell transformation sequence 2 mediates hepatocellular carcinoma cell epithelial-mesenchymal transition and promotes invasion and metastasis

目的 观察上皮细胞转化序列2（ECT2）对肝癌细胞增殖、迁移及侵袭能力及其对肝癌细胞上皮-间充质转化（EMT）的影响.方法 应用Western blot检测5例肝细胞癌标本及其配对癌旁组织ECT2的表达;通过慢病毒介导的方法构建过表达ECT2肝癌细胞,采用克隆形成实验和Transwell小室等方法观察其与对照组比较增殖、迁移及侵袭能力的变化,分析ECT2诱导肝癌细胞上皮-间充质转化进而促进转移复发的相关机制.结果 ECT2在肝癌组织中的表达较其在癌旁组织中明显升高;体外实验结果显示慢病毒转染的高表达ECT2的HEP3B细胞增殖较其对照组明显增高[（84.2±12.3）个比（26.7±5.7）个,P＜0.05],对照组及过表达组发生迁移的细胞数分别为[（38.6±15.6）个比（103.9±21.2）个,P＜0.05].通过上调肝癌细胞ECT2蛋白的表达能够诱导肝癌细胞发生EMT,其上皮化表型E-钙黏蛋白（E-cadherin）、紧密连接蛋白-1（ZO-1）的表达下调,间质化表型N-钙黏蛋白（N-cadherin）和纤维连接蛋白（Fibronectin）上调.结论 ECT2可能通过诱导肝癌细胞发生EMT进而参与调控其侵袭转移的进程。

Objective To investigate the relationship between the expression of epithelial cell transformation sequence 2 （ECT2） and epithelial-mesenchymal transition （EMT） in hepatocellular carcinoma cells.Methods The expression of ECT2 was detected by Western blotting in 5 cases of hepatocellular carcinoma （HCC） and the HCC adjacent tissue;After upregulation of ECT2 eppression in HEP3B cells using lentivirus transduction,the ability of proliferation and invasion was evaluated by transwell and colony formation assay.Results The expression of ECT2 was higher in HCC tumor tissue than adjacent tissue （P 〈 0.05）,The ECT2 upregulation significantly elevated the ability of HEP3B cells proliferation and invasion.Clone formation experiment results showed ECT2 over expression cell clone formation number was significantly higher than thatof control group 84.2 ± 12.3 vs.26.7 ± 5.7 （P 〈 0.05）.The number of cell migration occurred of over expression of ECT2 group and control group were 103.9 ± 21.2 vs.38.6 ± 15.6 （P 〈 0.05）.The characteristics of EMT were induced by lentivirus transduction of ECT2 into HEP3B cells,including the down-regulation of E-cadherin,and ZO-1,and the concomitant up-regulation of N-cadherin and Fibronectin.Conclusion ECT2 could mediate hepatocellular carcinoma cell EMT and promote invasion and metastasis.