三叶青黄酮对肺癌A549细胞生长抑制与MAPKs通路关系的研究

Radix tetrastigma hemsleyani flavone suppresses human lung carcinoma A549 cell by regulating MAPKs pathway

目的探讨三叶青黄酮对人非小细胞肺癌A549细胞增殖抑制、凋亡作用及其机制。方法采用CCK-8检测三叶青黄酮对A549细胞增殖的抑制作用;荧光显微镜观察Hoechst 33258染色的细胞凋亡形态变化;流式细胞仪检测细胞凋亡率;Western blot法检测p-p38、p-ERK,p-JNK蛋白表达变化。结果体外实验CCK-8检测显示三叶青黄酮各浓度组作用不同时间点细胞的抑制率比较,差异有显著性;呈剂量、时间依赖性。荧光显微镜观察经Hoechst 33258染色的细胞凋亡形态变化,显示细胞减少,细胞核改变和凋亡小体形成。流式细胞仪检测结果显示细胞凋亡率呈明显的量-效关系。Western blot法检测表明三叶青黄酮可以上调A549细胞中p-p38和p-ERK蛋白表达,且与浓度呈正相关。结论三叶青黄酮对人非小细胞肺癌A549细胞具有明显抑制增殖,促进凋亡作用,其机制可能与激活p38MAPK途径和ERK途径有关。

Aim To study the effect of Radix tetrastig- ma hemsleyani flavone （RTHF） on the growth and ap- optosis of human lung carcinoma A549 cells as well as its mechanisms. Methods A549 cells were treated with RTHF at different concentrations for different time in vitro, and the proliferation of A549 cell was detected by cell counting Kit-8 assay. Hoechst 33258 methods and Flow cytometry were adopted to determine cell ap- optosis. Expression of apoptosis-related MAPKs signal pathway proteins was detected by Western blot. Re- suits CCK-8 showed that RTHF had obvious anti-pro- liferation effect on A549 cells in a dose- and time-de- pendent manner. Microscope found typical manifesta- tions of cell apoptosis, and flow cytometric testing resuits showed that the treatment of apoptosis was signifi- cantly higher than the proportion in the control group （P 〈 0.01 ）. The protein expression of p-p38 and p- ERK were up-regulated significantly and that of p-JNK was little change in RTHF group when compared with the control group. Conclusion RTHF exerts anti- growth and apoptosis activity against human lung canc- er A549 cells, which may be associated with the up- regulation of MAPKs pathway proteins, the p-p38 and p-ERK.