IL-6调节Twist的表达对肝癌细胞BEL-7402上皮-间质转化的影响

摘要

目的 探讨白细胞介素6(interleukin-6,IL-6)通过调节Twist的表达调控肝癌细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)及侵袭转移的相关机制.方法 用IL-6处理肝癌细胞BEL-7402,采用Transwell实验检测细胞迁移侵袭能力.采用Western blot和qRT-PCR技术检测低表达Twist对E-cadherin和vimentin蛋白表达水平的影响和mRNA表达的变化.结果 IL-6处理BEL-7402细胞后侵袭和迁移能力明显提高.相较于空白对照组(CON组)和RNA干扰阴性对照组(RNAi-NC组),在转染组RNAi-Twist细胞中,E-cadherin蛋白和mRNA的表达水平明显增加;vimentin的蛋白表达水平明显下调,而mRNA的表达水平差异无显著性.加入50 ng/ml IL-6孵育细胞24 h后,CON组和RNAi-NC组E-cadherin蛋白和mRNA表达均较未处理时明显减少,RNAi-Twist组减少的趋势不明显;vimentin蛋白表达均较未处理时明显升高,而RNAi-Twist组升高不明显,相应mRNA的表达水平差异均无显著性.结论 IL-6可提高肝癌细胞BEL-7402体外侵袭、迁移能力,促进EMT过程,其机制可能通过调节Twist的表达,从而诱导发生EMT和侵袭转移.%Purpose To investigate of the meehanism of interleukin-6 ( IL-6 ) on epithelial-mesenchymal transition through the regulation of Twist expression in human hepatoma cell line BEL-7402. Methods Pretreatment with IL-6, cell migration and invasion were detected by transwell chamber assays. The influence of loss of Twist expression on the expression of E-cadherin and vimentin mRNAs and proteins was evaluated by quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) and Western blot. Results By pretreatment with IL-6, the ability of cell migration and invasion was significantly increased. Compared to non-transfected BEL-7402 cells ( control group ) and those tranfected with RNAi-NC ( RNAi negative control group ), the expression of E-cadherin mRNA and protein remarkably increased in BEL-7402 cells transfected with RNAi-Twist. Loss of Twist expression down-regulated vimentin protein expression, but had no significant impact on vimentin mRNA expression in BEL-7402 cells and those transfected with different siRNAs. After treated with IL-6 at a final concentration of 50 ng/ml for 24 h, the expression of E-cadherin mRNA and protein remarkably decreased in non-transfected BEL-7402 cells and those tranfected with RNAi-NC and the decreasing trend was not significant in BEL-7402 cells transfected with RNAi-Twist. The expression of vimentin protein remarkably increased in non-transfected BEL-7402 cells and those tranfected with RNAi-NC and the increasing trend was not significant in BEL-7402 cells transfected with RNAi-Twist. The expression of vimentin mRNA in BEL-7402 cells and those transfected with different siRNAs was not significantly different. Conclusion IL-6 could promote the cell invasion, migration and epithelial-mesenchymal transition process of human hepatoma cells by regulation of Twist expression and the expression of EMT protein ( E-cadherin and vimentin ).