IL-6在胃癌细胞株HGC-27上皮间质转化中的作用

Effects of IL-6 on epithelial-mesenchymal transition of HGC-27 cells in vitro

目的探讨白细胞介素-6(IL-6)对人胃癌细胞株HGC-27上皮间质转化(epithelial mesenchymal transition,EMT)的影响。方法体外用IL-6刺激培养HGC-27细胞,倒置显微镜下观察细胞形态学变化;实时荧光定量RT-PCR及western blot检测EMT相关标志分子E-cadherin、N-cadherin和Vimentin表达情况;Transwell迁移实验和平板克隆形成实验分别检测IL-6处理前后HGC-27细胞迁移和克隆形成能力的变化。结果 IL-6能诱导HGC-27细胞向间质细胞样形态转化,多数细胞表现为类似成纤维状,部分细胞呈明显的梭形;实时荧光定量RT-PCR结果表明,IL-6作用于HGC-27细胞后E-cadherin表达量明显降低(P<0.01),而N-cadherin和Vimentin表达则显著增强(P均<0.05)。western blot结果表明,IL-6作用HGC-27细胞后E-cadherin蛋白表达量降低(P<0.05),而N-cadherin和Vimentin蛋白表达量均增加(P均<0.05);Transwell实验及平板克隆形成实验结果显示,IL-6作用后HGC-27细胞的克隆形成能力及迁移能力均增强(P均<0.01)。结论 IL-6可诱导胃癌细胞发生EMT和进一步恶化的潜能。

Objective To investigate the effects of IL-6 on epithelial-mesenchymal transition（EMT） of human gastric adenocarcinoma cell line HGC-27 in vitro.Methods HGC-27 cells were cultured with DMEM medium containing IL-6（50 ng / mL）.The morphological changes of the cells were observed by invert phase contrast microscope.The expressions of EMT-associated genes,E-cadherin,Ncadherin and vimentin in HGC-27 cells were analyzed with real-time fluorescence quantitative PCR（qRT-PCR）,and the levels of Ecadherin,N-cadherin and vimentin were determined by western blotting.Transwell migration assay and plate clonality assay were conducted respectively to assess the changes of cell migration and colony-forming ability of HGC-27 cells with or without IL-6 treatment.Results HGC-27 cells acquired more typical spindle-shape phenotype of mesenchymal cells after co-cultured with IL-6.Most of the cells presented fibroid form.qRT-PCR showed that the levels of mesenchymal cell marker N-cadherin and vimentin in the HGC-27 cells treated with IL-6 increased significantly（P 0.05 respectively） and the level of E-cadherin was suppressed（P 0.01）.Western blotting analysis showed that the levels of N-cadherin and vimentin protein in HGC-27 cells upon the exposure to IL-6 increased significantly（P 0.05） while the level of E-cadherin protein was suppressed（P 0.05）.IL-6 also remarkably enhanced the migration and colony-forming ability of HGC-27 cells.Conclusion IL-6 may induce epithelial-mesenchymal transition of gastric cancer cells,which should be a potential contributing to tumor progress.