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目的:研究中药复方扶正祛邪方对肺癌A549细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)的抑制作用及机制研究。方法:采用MTT方法观察扶正祛邪方对人腺癌A549细胞的抑制作用;采用Transwell方法观察扶正祛邪方对人腺癌A549细胞侵袭的抑制作用;Western blot方法观察A549细胞EMT过程中上皮细胞、间质细胞标志物E-cadherin、Vimentin、FN的表达,转录因子Snail的蛋白表达,以及信号分子PSmad2/3的表达。结果:1MTT结果显示:扶正祛邪方对人腺癌A549细胞的抑制作用呈剂量和时间依赖。2Transwell侵袭实验结果显示:不同浓度的扶正祛邪方对人腺癌A549细胞的穿膜能力有抑制作用(P<0.05)。3Western blot结果显示:与模型组相比,扶正祛邪方上调E-cadherin蛋白表达,下调Vimentin、FN、P-Smad2/3蛋白表达(P<0.05);结论:1扶正祛邪方对A549细胞有直接的抑制作用;2扶正祛邪方对TGF-β1诱导人腺癌A549细胞EMT有抑制作用,其机制与抑制Smad2/3的磷酸化,进而减少Snail蛋白的表达有关。
Objective: To study the inhibitory effect and mechanism of Fuzheng Quxie Recipe on the epithelial-mesenchymal transition (EMT) in lung cancer A549 cells. Methods: MTT method was used to observe the inhibitory effect of Fuzhengquyu Decoction on human adenocarcinoma A549 cells. Transwell method was used to observe the inhibitory effect of Fuzhengquyu Decoction on human adenocarcinoma A549 cell invasion. Western blot was used to observe the effect of Fuzhengquyu Decoction , The expression of interstitial cell markers E-cadherin, Vimentin, FN, the transcription factor Snail, and the expression of PSmad2 / 3. Results: The results of 1 MTT showed that the inhibitory effect of Fuzheng Quxie prescription on human adenocarcinoma A549 cells was dose-and time-dependent. The results of 2Transwell invasion assay showed that different concentrations of Fuzhengqudefang had inhibitory effect on the transmembrane permeability of human adenocarcinoma A549 cells (P <0.05). 3Western blot results showed that Fuzhengqudefang up-regulated the expression of E-cadherin and down-regulated the expression of Vimentin, FN and P-Smad2 / 3 in the model group (P <0.05) Had a direct inhibitory effect; 2 Fuzhengquxue Fang on TGF-β1-induced human adenocarcinoma cell line A549 EMT inhibition of its mechanism and inhibition of Smad2 / 3 phosphorylation, thereby reducing the expression of Snail protein.