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目的:观察转化生长因子-β1(TGF-β1)诱导A549/DDP细胞发生上皮间质转化后的改变及补中益气汤含药血清的干预作用。方法:培养细胞株,将其分为D组(A549/DDP+空白血清)、DT组(A549/DDP+TGF-β1+空白血清)、DTB组(A549/DDP+TGF-β1+最佳浓度含药血清),采用MTT法鉴定A549/DDP细胞耐药性,普通显微镜下观察细胞形态变化,免疫细胞化学、Western Blot技术检测凋亡蛋白Fas、Fas L表达变化,TUNEL/DAPI荧光染色法检测早期细胞凋亡情况。结果:本实验中A549/DDP细胞株具有耐药性,且经TGF-β1诱导A549/DDP细胞后出现典型上皮间质转化形态,Fas、Fas L蛋白表达减少,细胞凋亡率下降(P<0.05,P<0.01)。补中益气汤含药血清干预后典型上皮间质转化形态消失,Fas、Fas L蛋白表达增加,细胞凋亡率上升(P<0.01)。结论:补中益气汤含药血清可能通过干预非小细胞肺癌耐药细胞上皮间质转化,影响Fas/Fas L信号通路促进细胞凋亡,改善其耐药性。
OBJECTIVE: To observe the changes of epithelial-mesenchymal transition induced by transforming growth factor-β1 (TGF-β1) in A549 / DDP cells and the effect of Buzhong Yiqi decoction-containing serum. Methods: The cell lines were cultured and divided into group D (A549 / DDP + blank serum), DT group (A549 / DDP + TGF-β1 + blank serum), DTB group ). The drug resistance of A549 / DDP cells was identified by MTT assay. The cell morphological changes were observed under a normal microscope. The expressions of Fas and Fas L proteins were detected by immunocytochemistry and Western Blot. The expressions of Fas and Fas L were detected by TUNEL / DAPI staining Death situation. Results: The A549 / DDP cell line was resistant in this experiment, and the typical epithelial-mesenchymal transitional morphology was found after A549 / DDP cells were induced by TGF-β1. The expression of Fas and Fas L decreased and the apoptosis rate decreased (P < 0.05, P <0.01). After the intervention of Buzhong Yiqi Decoction containing serum, the typical epithelial-mesenchymal transition disappeared, the expressions of Fas and Fas L increased, and the apoptosis rate increased (P <0.01). Conclusion: Buzhong Yiqi decoction-containing serum may affect the epithelial-mesenchymal transition of non-small cell lung cancer cells by affecting the Fas / Fas L signaling pathway to promote cell apoptosis and improve its drug resistance.