WWOX对肝内胆管细胞癌转移的影响

Effects of WWOX on metastasis of intrahepatic cholangiocarcinoma

目的 :检测包含WW域的氧化还原酶(WW domain containing oxidoreductase,WWOX)在肝内胆管癌(intrahepatic cholangiocarcinoma,ICC)组织及细胞的表达情况,分析其表达水平与ICC临床特征的相关性,并探索其对ICC细胞迁移、侵袭能力的影响。方法:采用半定量/定量PCR测定ICC组织及细胞中WWOX的表达水平;转染重组质粒pc DNA3.1B-WWOX-Flag,过表达ICC细胞内的WWOX水平;采用细胞划痕及侵袭实验,观察过表达WWOX对ICC迁移、侵袭能力的影响;蛋白免疫印迹检测迁移、侵袭通路相关分子。结果:WWOX在ICC组织及细胞中表达下调,其下调与ICC的转移明显相关(P<0.05);转染pc DNA3.1B-WWOX-Flag质粒可上调ICC细胞WWOX蛋白的表达;过表达WWOX抑制ICC细胞迁移、侵袭能力(P<0.001);过表达WWOX可激活细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)磷酸化,进而下调基质金属蛋白酶9的活性,最终抑制ICC的转移过程。结论:WWOX在ICC中表达下调,体外实验证实过表达WWOX抑制ICC的迁移、侵袭能力。

Objective：To detect the expression of WW domain containing oxidoreductase（WWOX） in intrahepatic cholangiocarcinoma（ICC） tissues and cells,to analyze the correlation between WWOX expression and clinical features of ICC,and to investigate its effect on cellular migration and invasion of ICC. Methods：The expression level of WWOX in ICC tissues and cell lines were measured by semi-quantitative real-time PCR and quantitative real-time PCR. The expression of WWOX in ICC cells was overexpressed by recombinant plasmid pc DNA3.1B-WWOX-Flag transfection. The effect of WWOX overexpression on migration and invasion of ICC was performed by wound healing and matrigel invasion assays. The potential signaling molecules involved in migration and invasion was measured by Western blot assay. Results：WWOX was down-regulated in ICC tissues and cell lines,and the downregulated WWOX was significantly associated with the metastasis of ICC（P〈0.05）. The expression of WWOX was upregulated by pc DNA3.1B-WWOX-Flag transfection in ICC cells. Overexpression of WWOX inhibited cellular migration and invasion of ICC（P〈0.001）. The overexpression of WWOX stimulated the activation of extracellular regulated protein kinases 1 / 2（ERK1 / 2）phosphorylation,which in turn reduced the activity of matrix metalloproteinases 9,resulting in metastasis suppression of ICC.Conclusion：WWOX was down-regulated in ICC,and overexpression of WWOX was proved to inhibit the ability of migration and invasion of ICC cells in vitro.