索拉非尼对食管癌EC9706细胞的生长抑制作用

目的 探讨索拉非尼（sorafenib）体外对EC9706细胞生长抑制作用及其机制。方法 qRTPCR检测sorafenib作用后MMP-2、MMP-9、TIMP1、AKT和Bcl-2 mRNA的表达；Western blot检测sorafenib作用后AKT、p-AKT、Bcl-2、MMP-2和TIMP1蛋白表达水平的变化；Hoechst/PI 染色荧光显微镜观察sorafenib诱导的细胞凋亡/坏死；细胞划痕实验观察sorafenib对EC9706细胞的迁移抑制作用。结果 与对照组相比，qRT-PCR显示sorafenib下调MMP-2、MMP-9、AKT、Bcl-2 mRNA表达，上调TIMP1 mRNA 表达（P<0.05）；Western blot显示sorafenib降低MMP-2、AKT、p-AKT、Bcl-2蛋白的表达水平，上调TIMP1蛋白表达水平（P<0.05）；荧光显微镜观察发现sorafenib诱导红染细胞增加（P<0.05）；细胞划痕实验发现sorafenib作用后划痕明显变宽，并且具有浓度依赖性（P<0.05）。结论 索拉非尼能够上调TIMP1表达水平、下调MMP-2、MMP-9、AKT、Bcl-2表达水平，可以抑制食管鳞癌EC9706细胞的迁移、促进凋亡坏死，这些可能是其产生抗肿瘤作用的机制之一。

Inhibitory Effect of Sorafenib on Growth of Human Esophageal Cancer Cells EC9706

Objective To investigate the inhibitory effect of sorafenib on the growth of esophageal cancer cells EC9706 in vitro and its mechanism. Methods Effect of sorafenib on the expression of MMP-2, MMP-9, TIMP1, AKT and Bcl-2 were detected by qRT-PCR. Effect of sorafenib on the expression of AKT, p-AKT, Bcl-2, MMP-2 and TIMP1 were detected by Western blot. Cell apoptosis/necrosis induced by sorafenib were observed by Hoechst/PI staining. The inhibitory effect of sorafenib on EC9706 cells migration was observed by cell scratch test. Results qRT-PCR data showed downregulation of MMP-2, MMP-9, AKT, Bcl-2 and upregulation of TIMP1 mRNA, compared with the control group(P<0.05); furthermore, Western blot analysis revealed the expression of MMP-2, AKT, p-AKT, Bcl-2 proteins were downregulated and TIMP1 protein expression was upregulated in the sorafenib groups compared with the control group(P<0.05).Sorafenib was found not only to inhibit the migration of EC9706 cells in vitro by cell scratch test, but also to promote cells apoptosis in vitro by Hoechst/PI apoptosis/necrosis detection(P<0.05). Conclusion Sorafenib could upregulate the expression of TIMP1, downregulate the expression of MMP-9, AKT, MMP-2 and Bcl-2,and inhibit the migration and promote apoptosis/necrosis of esophageal cancer cells EC9706. These may be its anticancer mechanism.