论文部分内容阅读
目的:观察益气除痰方是否能通过干预TGF-β信号途径,逆转肺癌A549细胞上皮间质转化(EMT)。方法:通过暴露于TGF-β_1环境中,诱导肺腺癌细胞株A549发生EMT,并以益气除痰方含药血清进行干预,相差显微镜下观察各组细胞形态变化;划痕实验、Transwell实验检测各组细胞迁移及侵袭能力;Western-blot检测各组细胞EMT相关蛋白及TGF-β信号通路蛋白Smad2/3的表达。结果:镜下可见经TGF-β_1孵育的A549细胞呈现明显的间质细胞形态,益气除痰方含药血清作用24 h后细胞A549细胞出现凋亡且间质化程度较低;划痕实验结果显示:与空白血清组比较,TGF-β_1+空白血清组A549细胞迁移及侵袭能力显著升高,益气除痰方可降低TGF-β_1所升高的细胞迁移及侵袭能力;Western-blot检测显示TGF-β_1可以激活Smad2/3通路下调E-cadherin的表达,上调Vimentin的表达,而益气除痰方含药血清可抑制该过程。结论:益气除痰方可能通过干预TGF-β/Smad信号通路上调E-cadherin表达,下调Vimentin表达,从而逆转肺癌A549细胞上皮间质转化,抑制肿瘤细胞的侵袭迁移。
Objective: To observe whether Yiqi Chu Tan Recipe can reverse the epithelial-mesenchymal transition (EMT) of lung cancer A549 cells by interfering with TGF-β signaling pathway. Methods: EMT was induced in lung adenocarcinoma cell line A549 by exposing to TGF-β 1, and intervention with Yiqi Chu Tan Decoction-containing serum. The morphology of the cells was observed under phase contrast microscope. The scratch test, Transwell test The migration and invasion ability of cells in each group were detected. The expression of EMT-related protein and TGF-β signaling pathway protein Smad2 / 3 were detected by Western-blot. Results: The A549 cells cultured with TGF-β 1 showed obvious interstitial morphology under the microscope. A549 cells were apoptosis and the degree of interstitialis was lower after 24 h treatment with Yiqi Zutan Recipe containing serum. Scratch experiment The results showed that compared with the blank serum group, the migration and invasion ability of A549 cells in TGF-β 1 + blank serum group was significantly increased, and Yiqi Chu Tan could reduce the cell migration and invasion ability induced by TGF-β 1; Western-blot showed TGF-β 1 can activate Smad2 / 3 pathway to down-regulate the expression of E-cadherin and up-regulate Vimentin expression. Conclusion: Yiqi Zutan Recipe can up-regulate the expression of E-cadherin and down-regulate the expression of Vimentin in TGF-β / Smad signaling pathway, thereby reversing the epithelial-mesenchymal transition of lung cancer A549 cells and inhibiting the invasion and migration of tumor cells.