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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 investigate the effect of COE on invasion and metastasis of human gastric cancer cell line SGC-7901 and its molecular mechanism. Methods The effect of COE on the proliferation of human gastric cancer SGC-7901 cells was detected by MTT assay. The effect of COE on the invasion and metastasis of SGC-7901 cells was detected by Transwell chamber assay. The protein levels of MMP2, MMP9 and TIMP1, TIMP3 in SGC-7901 cells treated with COE were detected by Western blotting. The mRNA expression of MMP2, MMP9, TIMP1 and TIMP3 in SGC-7901 cells treated with COE was detected by RT-PCR. Results The results of Transwell assay showed that the number of transmembrane cells in SGC-7901 cells treated with 20, 40 and 80 mg / L of COE decreased significantly and in a concentration-dependent manner. Western blotting showed that the expression of MMP2 and MMP9 in SGC-7901 cells treated with 20, 40 and 80 mg / L of COE was significantly decreased, while the expressions of TIMP1 and TIMP3 were significantly up-regulated. RT-PCR results showed that the m RNA levels of MMP2 and MMP9 increased slightly in 20, 40 and 80 mg / L COE-treated SGC-7901 cells while the levels of m RNA in TIMP1 and TIMP3 were significantly increased (P <0.001). Conclusion COE can significantly inhibit the invasion and migration of human gastric cancer cell line SGC-7901, and its mechanism may be directly related to the down-regulation of MMP2 and MMP9 protein levels. The realization of its function is likely to be achieved by directly up-regulating TIMP1 and TIMP3 m RNA transcription levels and thus increasing TIMPs protein expression levels.