南方红豆杉水提物诱导人肺癌A549细胞凋亡及其分子机制的实验研究

Experimental Study on Apoptosis Induced by Aqueous Extract of Taxus chinensis in Human Pulmonary Carcinoma Cell A549 and Its Molecular Mechanisms

目的探讨南方红豆杉水提物诱导人肺癌A549细胞的凋亡作用及其分子机制。方法采用噻唑蓝(MTT)法检测南方红豆杉水提物对A549细胞的增殖抑制作用;以光镜、电镜、膜联蛋白Ⅴ-FITCPI(An-nexinⅤ-FITC/PI)标记法检测细胞凋亡;以蛋白印迹法分析南方红豆杉水提物对细胞外信号调节蛋白激酶1/2(ERK1/2),c-jun氨基未端激酶1/2(JNK1/2)和生存素(survivin)蛋白表达的影响。结果南方红豆杉水提物能抑制A549细胞的增殖;南方红豆杉水提物处理A549细胞后,光镜与电镜下可见到明显的凋亡细胞;Annexin Ⅴ-FITC/PI标记法的定量检测进一步证实了南方红豆杉水提物诱导细胞凋亡的作用;蛋白印迹检测表明南方红豆杉水提物能使survivin表达下降,对ERK1/2和JNK1/2表达无影响。结论南方红豆杉水提物能显著抑制A549细胞增殖,诱导细胞凋亡,主要通过调节survivin蛋白的表达来实现。

Objective To investigate the apoptosis inducing effects of aqueous extract of Taxus chinensis（AETC） in human lung cancer cell A549 and its molecular mechanisms.Methods Using MTT assay,the inhibition of AETC on the proliferation of A549 cells was detected.The apoptosis was detected by light and electron microscopy,and Annexin Ⅴ-FITC/PI staining.The expressions of ERK1/2,JNK1/2,and survivin were analyzed by Western blot.Results AETC could inhibit the proliferation of A549 cells.Obvious apoptotic cells could be seen under light and electron microscope in AETC treated A549 cells.AETC＇s induction on the cell apoptosis was further confirmed by Annexin Ⅴ-FITC/PI labeled quantitative detection.Western blot assay showed that AETC could decrease the survivin expressions.AETC showed no effects on ERK1/2 or JNK1/2 protein expressions.Conclusion AETC could significantly inhibit the proliferation of A549 cells and induce the apoptosis,which was mainly achieved through regulating the expressions of survivin.