法舒地尔对胃癌MGC-803细胞增殖、侵袭和迁移的影响

摘要

Objective To investigate the effect of Fasudil on the pfoliferation,invasion and migration of gastric cancer MGC-803 cells and their probable mechanisms. Methods MGC-803 cells were treated with different concentration of Fasudil and cell num-ber was counted. And the subsequent experiments were conducted in 3 groups respectively contained 10,25,50 μmol/L of Fasudil, the MGC-803 cells treated with Fasudil were taken as the treatment group,while those not treated with Fasudil were taken as the control group. The cells were cultured for 4 days(0,24,48,72,96 h),and the cell growth folds were detected by CCK-8.The ability of cell migration was detected by wound-healing assay. The ability of cell invasion was detected by Transwell. The protein expression levels of vascular endothelial growth factor (VEGF),matrix metalloproteinases 9(MMP-9),matrix metalloproteinases14 (MMP-14)and epi-thelial mesenchymal transformation (EMT)markers Vimentin and E-cadherin were detected by Western blotting. Results The toxicity test showed that Fasudil inhibited the cell growth of MGC-803 cells in a dose-dependent manner and the cell growth folds were reduced in the treatment group. There was an obvious decrease in the growth folds compared to the control group (P＜0.01). The results of wound healing assay showed that the wound closure rate in the treatment group was lower than that of the control group,and in a dose-dependent manner(P＜0.01). Transwell results showed that the number of invasive cells in the treatment group was decreased signifi-cantly compared with the control group (P＜0.01). Besides,Fasudil significantly decreased the expression levels of VEGF,MMP-9, MMP-14 and Vimentin,and up-regulated the expression levels of E-cadherin. Conclusion Fasudil may inhibit cell proliferation,mi-gration and invasion of MGC-803 cells,reduce the activity of MMP-9,MMP-14 and VEGF,which will provide new sight and theoreti-cal basis for the treatment of gastric cancer.%目的 探讨法舒地尔对胃癌MGC-803细胞增殖、侵袭和迁移的影响及可能机制.方法 采用不同浓度的法舒地尔处理胃癌MGC-803细胞,检测细胞数目;选择无显著细胞毒性的3组浓度(10、25、50 μmol/L)法舒地尔进行后续实验,将经药物处理的胃癌MGC-803细胞作为处理组,而未经药物处理的胃癌MGC-803细胞作为对照组;细胞培养0、24、48、72、96 h后,CCK-8法检测细胞增殖倍数;划痕实验检测细胞迁移能力;Transwell法检测细胞侵袭能力;Western blotting检测细胞中血管内皮细胞生长因子(VEGF)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶14(MMP-14)和上皮间质转化(EMT)的标志蛋白波形蛋白(Vimentin)和上皮型钙黏蛋白(E-cadherin)的表达水平.结果 CCK-8法检测结果显示,法舒地尔以剂量依赖性的方式抑制胃癌MGC-803细胞的生长,法舒地尔浓度越高,处理组的细胞数量越少,增殖倍数明显降低(P＜0.01);划痕实验结果表明,处理组伤痕愈合率明显较对照组降低,且法舒地尔浓度越高,处理组伤痕愈合率越低(P＜0.01);Transwell结果显示,与对照组比较,处理组侵袭的细胞数目明显减少(P＜0.01);与对照组比较,法舒地尔明显下调VEGF、MMP-9、MMP-14和Vim-entin蛋白的表达水平,而上调E-cadherin蛋白的表达水平.结论 法舒地尔可抑制胃癌MGC-803细胞的的增殖、迁移和侵袭能力,降低MMP-9、MMP-14、VEGF表达水平,为胃癌的治疗研究提供新思路和理论基础.