软坚散结颗粒对人胃腺癌SGC-7901细胞EMT的抑制作用及相关机制

目的:上皮间质转化(epithelial-mesenchymal transition,EMT)现象的发生发展是肿瘤转移的关键因素之一,对胃癌EMT现象的研究可能有助于控制胃癌转移。本课题旨在研究软坚散结方(RJSJ)对人胃腺癌SGC-7901细胞EMT的抑制作用并探讨相关机制。方法:采用噻唑蓝(MTT)比色法观察RJSJ对SGC-7901细胞的抑制作用;采用transwell方法观察RJSJ对SGC-7901细胞侵袭的抑制作用;采用实时荧光定量PCR(Real-time PCR)和蛋白免疫印迹法(Western blot)观测SGC-7901细胞EMT过程中锌指E盒结合同源盒蛋白1(Zinc finger E-box binding homeobox 1,Zeb1),锌指E盒结合同源盒蛋白2(Zinc finger E-box binding homeobox 2,Zeb2)以及E-钙黏蛋白(E-cadherin)mRNA和蛋白的表达。结果:(1)MTT结果显示,不同浓度RJSJ组均能有效抑制SGC-7901细胞的增殖(P0.05),RJSJ对SGC-7901细胞的抑制作用呈剂量和时间依赖。(2)transwell侵袭实验结果显示,不同质量浓度RJSJ(250,500,1 000 mg·L-1)对SGC-7901细胞的穿膜能力有不同程度的抑制作用(P0.05)。(3)Real-time PCR和Western blot结果显示,与空白组比较,RJSJ各组均能使E-cadherin mRNA和蛋白表达增加,Zeb1,Zeb2mRNA和蛋白表达减少,其中RJSJ高剂量组能够明显上调E-cadherin mRNA和蛋白表达,下调Zeb1,Zeb2 mRNA和蛋白表达(P0.05)。结论:RJSJ对SGC-7901细胞有直接的抑制作用;RJSJ对SGC-7901细胞EMT有抑制作用,其机制可能与下调Zeb1,Zeb2 mRNA及蛋白表达,上调E-cadherin mRNA及蛋白表达有关。

Intervention Effect and Mechanism of Ruanjian Sanjie Granules for Gastric Carcinoma Cell SGC-7901

Objective: The development of epithelial-mesenchymal transition (EMT) is one of the key factors for tumor metastasis, so the research on the phenomenon of EMT may be conducive to control gastric cancer metastasis. This paper is aimed to explore the intervention effect and mechanism of Ruanjian Sanjie (RJSJ) granules for the invasion and metastasis of gastric carcinoma SGC-7901 cells.Method: Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay was used to observe the inhibitory effect of RJSJ on SGC-7901 cells proliferation; transwell assay was used to explore the inhibitory effect of RJSJ granules on the invasion of gastric carcinoma SGC-7901 cells; the mRNA and protein expression levels of E-cadherin, Zinc finger E-box binding homeobox 1 (Zeb1) and Zinc finger E-box binding homeobox 2 (Zeb2) were examined by Real-time PCR and Western blot respectively.Result: MTT experimental results showed that all different concentrations of RJSJ granules could inhibit the proliferation of gastric carcinoma SGC-7901 cells (P<0.05), and the inhibitory effect was in a dose and time dependent manner. transwell assay showed that RJSJ granules (concentrations of 250, 500, 1 000 mg · L^-1 respectively) could inhibit invasion of gastric carcinoma SGC-7901 cells(P<0.05). Real-time PCR showed that RJSJ granules could up-regulate the mRNA expression levels of E-cadherin and down-regulate the mRNA expression levels of Zeb1 and Zeb2; while Western blot showed that RJSJ granules could up-regulate the protein expression levels of E-cadherin and down-regulate the protein expression levels of Zeb1 and Zeb2(P<0.05).Conclusion: RJSJ granules could inhibit SGC-7901 cells proliferation; RJSJ granules had an inhibitory effect on SGC-7901 cells EMT, and the mechanism may be associated with down-regulating Zeb1 and Zeb2 mRNA and protein expression and up-regulating E-cadherin mRNA and protein expression.