摘要
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：Human adenocarcinoma A549 cells for the study,to explore the intervention effect and mechanism research of Fuzhengquxie Decoction in the invasion and metastasis of lung cancer.Methods：MTT assay was used to determine A549 cells proliferation.The expression of E-cadherin、N-cadherin and P-Smad2 /3 were examined by Western blot; TGF-β3mRNA was examined by RT-PCR.Results：MTT experimental results show that the inhibitory effect of different concentrations of compound on human adenocarcinoma A549 cells,with the increase of time,the increase of concentration,the stronger the inhibition.Western Blot showed that Fuzhengquxie Decoction could up-regulated the expression of E-cadherin protein,down-regulated the expression of N-cadherin and P-Smad2 /3 protein respectively; RT-PCR showed that Fuzhengquxie Decoction could down-regulated the expression of TGF-β3mRNA.Conclusion：Fuzhengquxie Decoction could inhibit A549 cells proliferation;Fuzhengquxie Decoction probably inhibited the migration and invasion of human adenocarinoma A549 cells via regulation of TGF-β3mRNA expression.
出处
《辽宁中医杂志》
CAS
北大核心
2015年第4期873-875,共3页
Liaoning Journal of Traditional Chinese Medicine
基金
上海市自然科学基金(09ZR1429800)
国家中医药管理局"十二五"重点专科项目基金(ZJ0901ZL020)
