消癌平注射液对人肝癌HepG2和MHCC97H细胞侵袭和转移抑制作用及分子机制研究

Effects of Xiaoaiping Injection on Cell Migration and Invasion of Human Cell Lines HepG2 and MHCC97H and the Molecular Mechanism

目的:研究消癌平注射液(XAP)对人肝癌HepG2和MHCC97H细胞侵袭和转移抑制作用及分子机制。方法:MTT法检测XAP对人肝癌细胞株HepG2和MHCC97H的增殖抑制作用,通过划痕实验、Transwell实验检测XAP对HepG2和MHCC97H细胞迁移能力的抑制作用,采用Transwell包被基质胶实验检测XAP对HepG2和MHCC97H细胞侵袭能力的抑制作用,应用Western blotting方法检测XAP对HepG2和MHCC97H细胞MYOF、MMP-2、MMP-9蛋白表达的影响。结果:MTT法结果显示,当XAP浓度为80μl·ml^(-1)作用24 h时,HepG2和MHCC97H细胞的存活率约为70%; XAP浓度为120μl·ml^(-1)及160μl·ml^(-1)时,对HepG2和MHCC97H细胞的增殖抑制作用逐渐增强(P<0.01)。48 h时,XAP呈剂量依赖性抑制HepG2和MHCC97H细胞的增殖(P<0.05或P<0.01)。划痕实验和Transwell实验结果显示XAP呈剂量依赖性地抑制佛波酯(12-O-tetradecanoylphorbol13-acetate,TPA)诱导的HepG2和MHCC97H细胞迁移(P<0.05或P<0.01)。Transwell包被基质胶实验显示XAP呈剂量依赖性地抑制TPA诱导的HepG2和MHCC97H细胞的侵袭(P<0.05或P<0.01)。Western blotting结果表明XAP抑制TPA诱导的HepG2和MHCC97H细胞中MYOF、MMP-2、MMP-9蛋白表达的升高(P<0.05或P<0.01)。结论:消癌平注射液能够抑制肝癌HepG2和MHCC97H细胞的侵袭和转移,其抑制作用可能与调控MYOF从而抑制基质金属蛋白酶MMP-2、MMP-9表达相关。

Objective: To investigate the inhibitory effect and the Molecular mechanism of Xiaoaiping injection (XAP) in human hematoma cell lines HepG2 and MHCC97H. Methods: In vitro anti-proliferation activity of XAP on human hematoma cell lines HepG2 and MHCC97H were investigated using MTT assay. The cells were treated with different concentration (40,80,120,160 μl·ml-1),migration and invasion ability were detected by wound healing and transwell chamber assay. Western blotting was used to detect the protein expression of MYOF,MMP-2,MMP-9 in the cells HepG2 and MHCC97H. Results: After treated with XAP,the proliferation on HepG2 and MHCC97H cells were inhibited gradually.These experiment groups were compared with those in control group,the difference was statistically signicant (P<0.05 or P<0.01). The results of wound healing and transwell chamber assays showed that the cells numbers of migration and invasion were decreased with the elevated concentration of XAP,compared with those in TPA group,the difference was statistically significant (P<0.05 or P<0.01). Western blotting results showed that the expression of MYOF,MMP-2 and MMP-9 was decreased obviously with the elevated concentration of XAP,compared with those in TPA group,the difference was statistically significant (P<0.05).Conclusion: XAP can effectively inhibit the migration and invasion ability of human hematoma cell lines HepG2 and MHCC97H. One of the mechanisms is perhaps the down-regulation of the MMP-2 and MMP-9 through suppressing the MYOF.