高尔基体膜蛋白Ⅱ在肝细胞肝癌转移、复发和上皮-间充质转化的相关机制

Golgi phosphoprotein 2 as a promoter of epithelial to mesenchymal transition in hepatocellular carcinoma

目的探讨高尔基体膜蛋白Ⅱ（GOLPH2）促进人肝细胞肝癌（HCC）细胞上皮-间充质转化（EMT）转移复发的机制。方法利用反转录-聚合酶链反应（RT—PCR）检测8种肝癌细胞株中C01，PH2的表达，选择其中高表达GOLPH2的MHCC．97H肝癌细胞利用慢病毒载体介导的RNA干扰GOLPH2的表达，观察侵袭、转移能力及EMT相关生物学特性是否因GOLPH2表达水平改变而发乍变化。结果RT—PCR结果显示GOLPH2的表达在HCC细胞株中随着恶性程度的增加而明显上升。在体外实验显示GOLPH2-RNA干扰（RNAi）转染组细胞克隆形成数（17．8±7．5）个显著低于对照组（41．7±5．3）个（P〈0．05）。转染组及对照组发生迁移的细胞数分别为（27．84-11．6）个比（278．2±23．4）个（P〈0．01）。转染组的迁移和侵袭能力明显减弱。同时转染组细胞间质表型N-钙黏蛋白（N—cadherin）和波形蛋白（Vimentin）下调而上皮表型E-钙黏蛋白（E—cadherin）、紧密连接蛋门-1（ZO-1）的表达上调（P〈0．01）。结论在HCC细胞中GLOPH2可通过调节EMT进而调控肝癌细胞侵袭转移。

Objective To explore the roles of golgi phosphoprotein 2 （GOLPH2） in hepatocellular carcinoma （HCC） metastatic progression as a promoter of epithelial to mesenchymal transition （EMT）. Methods We evaluated the expression of GOLPH2 in 8 HCC cell lines using reverse transcriptase - poly- merase chain reaction （RT- PCR）. Lentiviral -mediated RNA interference was used to obtain a stable HCC cell line with low expression of GLOPH2. In vitro we also investigated the mechanisms of GLOPH2 in EMT metastasis in HCC cell lines. Results RT - PCR showed that the mRNA levels of GLOPH2 in high - metaststic HCC cell lines were significantly increased in comparison to low - metaststic ones. Clone forma- tion experiment results showed clone formation number of RNAi transfection cells （ 17.8 ± 7.5 ） was signifi- cantly lower than that （ 41.7 ±5.3 ） of control group （ P 〈 O. 05 ）. The number of migrating cells in low GOLPH2 expression group and control group was 27. 8 ± 11.6 and 278. 2 ± 23.4 respectively. In vitro, the characteristics of EMT were induced by lentivirus transduction of GOLPH2 into MHCC -97H cells, inclu- ding the up - regulation of E - cadherin and ZO - 1, and the concomitant down - regulation of N - cadherin and Vimentin. Conclusion Our findings have uncovered a novel role for GOLPH2 in HCC metastasis, and it may become the molecular targets for intervention of tumor metastasis.