YM-155协同顺铂杀伤非小细胞肺癌细胞的机制研究

Mechanism of Synergistic Effect of YM-155 on Cisplantin-Induced Cytotoxicity Against NSCLC

关键词:  YM-155  survivin  顺铂  非小细胞肺癌  凋亡

Key words:  YM-155  survivin  cisplatin  NSCLC  apoptosis

摘要:

目的 探讨survivin小分子抑制剂YM-155对非小细胞肺癌顺铂化疗的协同治疗作用并研究其机制。方法 MTT法检测YM-155和顺铂对非小细胞肺癌细胞系A549的杀伤活性。Western blot试验检测A549细胞中survivin、细胞色素C、凋亡诱导因子、caspase-9和caspase-3的表达水平。流式细胞术检测A549细胞的线粒体膜电位和凋亡率。结果 YM-155能增强顺铂的抗肺癌活性，显著降低顺铂对A549细胞的半抑制浓度（IC₅₀）。YM-155处理能显著抑制A549细胞中survivin的表达。顺铂+YM-155组A549细胞的线粒体膜电位明显低于顺铂单治疗组。顺铂+YM-155组A549细胞的凋亡率，细胞色素C、凋亡诱导因子释放水平以及caspase-9、caspase-3活化水平均明显高于顺铂单治疗组。顺铂+YM-155+survivin质粒组A549细胞的相对细胞活力和线粒体膜电位明显高于顺铂+YM-155组。顺铂+YM-155+survivin质粒组A549细胞的凋亡率、细胞色素C、凋亡诱导因子释放水平以及caspase-9、caspase-3活化水平均明显低于顺铂+YM-155组。结论 YM-155通过抑制survivin的表达能够增强顺铂对非小细胞肺癌细胞线粒体途径凋亡的诱导活性。

Abstract:

OBJECTIVE To explore the adjuvant effect of YM-155 on cisplatin-based chemotherapy in non-small cell lung cancer(NSCLC). METHODS MTT assay was performed to evaluate the cytotoxicity of YM-155 and cisplatin to A549 cells. Western blot analysis was conducted to detect the expression of survivin, cytochrome C, apoptosis inducing factor, caspase-9 and caspase-3 in A549 cells. Flow cytometry analysis was performed to detect the mitochondrial membrane potential and apoptosis of A549. RESULTS YM-155 conled enhance the anti-lung cancer activity of cisplation, and decreased the IC₅₀ of cisplatin to A549. YM-155 inhibited the expression of survivin in A549. Mitochondrial membrane potential of A549 in cisplatin+YM-155 group was significantly lower than that in the cisplatin single treatment group. Apoptotic rate, release of cytochrome C and apoptosis inducing factor, activation of caspase-9 and caspase-3 in the cisplatin+YM-155 group was obviously higher than these in the cisplatin single treatment group. On the other hand, relative cell viability and mitochondrial membrane potential of A549 in cisplatin+YM-155+survivin plasmid group was significantly higher than those in the cisplatin+YM-155 group. However, apoptotic rate, release of cytochrome C and apoptosis inducing factor, activation of caspase-9 and caspase-3 in the cisplatin+YM-155+survivin plasmid group was obviously lower than those in the cisplatin+YM-155 group. CONCLUSION YM-155 enhances the cisplatin-induced mitochondrial apoptosis in NSCLC cells through inhibition of survivin.