南蛇藤提取物通过调控基质金属蛋白酶组及其抑制因子抑制人胃癌SGC-7901细胞侵袭转移的研究

Research on effect of Celastrus orbiculatus extract on inhibiting invasion and metastasis of human gastric cancer SGC-7901 cells by mediating matrix metalloproteinases group and their inhibitors

目的探讨南蛇藤提取物(COE)对人胃癌SGC-7901细胞的侵袭和转移的影响及其分子机制。方法 MTT法检测COE对人胃癌SGC-7901细胞增殖的影响;Transwell小室实验检测COE对SGC-7901细胞侵袭和转移能力的影响。Western blotting检测经COE作用后的SGC-7901细胞中金属基质蛋白酶(MMPs)MMP2、MMP9和金属基质蛋白酶抑制因子(TIMPs)TIMP1、TIMP3的蛋白水平。RT-PCR检测经COE作用后的SGC-7901细胞中MMP2、MMP9、TIMP1、TIMP3的m RNA表达情况。结果 Transwell实验显示,经20、40、80 mg/L COE处理过的SGC-7901细胞穿膜数明显减少并呈一定的浓度依赖性。Western blotting结果显示,经20、40、80 mg/L COE处理过的SGC-7901细胞,MMP2、MMP9蛋白的表达水平明显降低,而TIMP1、TIMP3蛋白表达水平明显上调。RT-PCR结果显示,经20、40、80 mg/L COE处理过的SGC-7901细胞中MMP2和MMP9的m RNA水平有轻微升高趋势,而TIMP1和TIMP3的m RNA水平明显大幅上调(P<0.001)。结论 COE能明显抑制人胃癌SGC-7901细胞的侵袭和迁移,其机制可能与下调MMP2、MMP9蛋白水平直接相关。而其作用的实现,很可能是通过直接上调TIMP1和TIMP3的m RNA转录水平从而提高TIMPs蛋白表达水平实现的。

Objective To study the effect and the mechanism of Celastrus orbiculatus extract（COE） on inhibiting the invasion and metastasis of human gastric cancer SGC-7901 cells. Methods The groups of different cells were set up, such as negative control group, COE with different concentration groups, and positive control group. The effects of COE on the cytotoxicity and proliferation of human gastric cancer SGC-7901 cells were determined by MTT assay. The effect of COE on the metastatic ability of human gastric cancer SGC-7901 cells in vitro was observed by invasion assay and migration assay. The effects of COE on the expression of matrix metalloproteinases（MMPs） and their inhibitors in human gastric cancer SGC-7901 cells were examined by Western blotting. After the treatment in different groups, m RNAs of MMP2, MMP9, tissue inhibitors of metalloproteinases（TIMPs）, TIMP1, and TIMP3 in cells were detected by RT-PCR. Results Compared with the negative control group, human gastric cancer SGC-7901 cells treated with COE at different concentration exhibited a dose-dependent growth inhibition. COE obviously reduced the migration and invasion potential of human gastric cancer SGC-7901 cells. In addition, the expression levels of MMP2 and MMP9 were dramatically suppressed by COE in a concentration-dependent manner, while the expression levels of TIMP1 and TIMP3 were raised. RT-PCR showed that m RNAs of TIMP1 and TIMP3 were raised with the increase of drug concentration. Conclusion COE could significantly inhibit the invasion and metastasis of human gastric cancer SGC-7901 cells via an up-regulation of the expression levels of TIMP1 and TIMP3 and down-regulation of the expression levels of MMP-2 and MMP-9.