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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;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、N-cadherin的表达,以及信号分子P-Smad2/3的表达;RT-PCR检测扶正祛邪方对I型胶原诱导人腺癌A549细胞EMT过程中TGF-β3mRNA的影响。结果:(1)MTT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。(2)Western 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N-cadherin、P-Smad2/3的蛋白表达(P<0.05);RT-PCR结果显示:扶正祛邪方下调TGF-β3mRNA的表达,呈浓度依赖性。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I型胶原诱导TGF-β3mRNA表达,进而抑制Smad2/3的磷酸化,上调E-cadherin蛋白表达,下调N-cadherin蛋白表达有关。
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. Western blot was used to observe the expression of E-cadherin and N-cadherin in epithelial cells and interstitial cells during EMT in A549 cells. The expression of P-Smad2 / 3 was detected by RT-PCR. The effect of Fuzhengqudejie Decoction on TGF-|Â3mRNA in EMT induced by type I collagen in human adenocarcinoma A549 cells was detected by RT-PCR. Results: (1) The results of MTT showed that the inhibitory effect of Fuzheng Quxie Fang on human adenocarcinoma A549 cells was dose-and time-dependent. (2) Compared with the model group, Fuzhengquyu recipe upregulated the protein expression of E-cadherin and down-regulated the protein expression of N-cadherin and P-Smad2 / 3 (P <0.05) Fuzhengqixie Fang down-regulate the expression of TGF-β3 mRNA in a concentration-dependent manner. Conclusion: (1) Fuzhengquyu Decoction has a direct inhibitory effect on A549 cells; (2) Fuzhengquyu Decoction can inhibit the EMT of TGF-β1-induced human adenocarcinoma A549 cells, and its mechanism and inhibition of type I collagen-induced TGF- β3mRNA expression, and then inhibit Smad2 / 3 phosphorylation, upregulation of E-cadherin protein expression, down-regulation of N-cadherin protein expression.