麦门冬合千金苇茎汤抑制A549细胞增殖作用及其机制

Mechanisms of Proliferative Inhibition by Maimendong & Qianjinweijing Decoction in A549 Cells

背景与目的采用抗肿瘤中药治疗恶性肿瘤是具有中国特色的肿瘤治疗方式。本研究旨在体外筛选出麦门冬合千金苇茎汤抑制人肺癌A549细胞增殖的有效部位,并对其进行分子机制探讨。方法通过MTT法和细胞克隆形成试验体外初步筛选出麦门冬合千金苇茎汤抑制A549细胞增殖的有效部位;流式细胞仪进行细胞周期分析;Hoechst33258染色观察凋亡形态的变化;Westernblot对肺癌相关通路进行检测。结果乙酸乙酯萃取部位可显著抑制肺癌A549细胞的生长,且对正常细胞HFL-1无明显影响;与对照组相比,10μg/mL乙酸乙酯萃取部位作用后克隆形成抑制率达73.86%(P<0.01),流式检测凋亡率为33.86%(P<0.01),荧光显微镜下可见凋亡形态学特征;抗凋亡蛋白EGFR和ERK表达显著下调(P<0.01)。结论麦门冬合千金苇茎汤乙酸乙酯萃取部位能显著抑制A549细胞的生长,并通过下调EGFR/ERK信号转导通路诱导细胞凋亡,因此应进一步分离,明确复方抗癌作用的物质基础。

Background and objective Traditional Chinese medicine is an approach for malignant tumor treatment with Chinese characteristics. The aim of this study is to investigate the inhibitory effects of Maimendong qianjinweijing decoction extract on A549 human lung cancer cell line proliferation and explored its probable molecular mechanisms. Methods A549 cells were treated with drugs in different does and time. The effects on the proliferation of A549 cells were detected by MTT assay and clonogenic assay in vitro. Cell cycle was analyzed by flow cytometry. Morphological changes of the apoptosis of cancer cells were observed by Hochest 33258 staining. Western blot was performed to detect apoptosis-related gene expression. Results Ethyl acetate extract inhibited the growth of A549 cells but not in HFL-1 cells. Compared with controls, administration of 10 μg/mL ethyl acetate extract resulted in 73.86% decrease in colony formation （P0.01）, apoptotic rates of 33.86% （P0.01）, and morphological changes of apoptosis in A549 cells. The expression of anti-apoptotic protein EGFR and ERK were significantly down-regulated （P0.01）. Conclusion Ethyl acetate extract might inhibit proliferation and induce apoptosis in A549 cells via downregulation of EGFR/ERK signal transduction pathway. Therefore, ethyl acetate extract should be further separated in order to identify the material fundamentals on anti-cancer effect.