吗啡对顺铂诱导的肺癌A549细胞的生物学影响

The biological impaction of morphine on lung cancer A549 cells induced by cisplatin

目的:探讨吗啡联合顺铂处理肺癌A549细胞后,细胞增殖能力和细胞凋亡水平的变化。方法:选择不同浓度吗啡处理肺癌A549细胞,CCK8法检测6、12、24和48 h时点的细胞增殖抑制率;再采用30μg/m L顺铂单药联合筛选出的最佳抑制率浓度的吗啡处理细胞,48 h后检测细胞增殖活力,同时采用Western blot电泳检测顺铂单药、吗啡,以及二者联合分别处理细胞后,凋亡相关蛋白Bax、Bcl-2、Caspase-3、survivin表达量的变化。结果:CCK8法显示500μg/m L浓度的吗啡单独处理肺癌细胞48 h时,能最明显抑制细胞的增殖(P<0.05),抑制率达到41.2%;与单纯使用顺铂抑制细胞增殖效果(47.21%)相比,顺铂30μg/m L联合500μg/m L吗啡对细胞的抑制率达77.45%,差异具有统计学意义(P<0.05);Western blot实验显示顺铂联合500μg/m L吗啡与单纯顺铂药物作用相比,Bcl-2、survivin蛋白表达量降低,Bax、Caspase-3蛋白的表达升高。结论:选择合适的吗啡浓度可加强顺铂对肺癌细胞的抑制增殖和促凋亡功能,这种对肺癌细胞的生物学效应可能与促进凋亡蛋白Bax、Caspase-3表达上调和凋亡抑制蛋白的下调相关。

Objective：To analyze the effect and mechanism of the proliferation inhibition and apoptosis in human lung carcinoma after using morphine combined with cisplatin chemotherapy. Methods：Human lung ad-enocarcinoma cell line A549 was cultured in conventional 1 640 medium. Cell proliferation activity for cisplatin alone or in combination with morphine after treatment for 6 h, 12 h, 24 h and 48 h were detected by CCK8. In addition, Western Blot electrophoresis was adopted to detect cisplatin alone or in combination with the optimal concentration of morphine after 48 h. Then changes of apoptosis related proteins Bax, Bcl-2 Caspase-3, survivin expression level were analyzed. Results：After 24 h, or 48 h, the concentration of 500μg/mL morphine in combi-nation with cisplatin group, compared to morphine and cisplatin alone group, could signiifcantly increase cancer cell proliferation inhibitory effect （P〈0.05）. While combined with low-dose concentrations of morphine-treated group, compared with cisplatin alone, the difference was not statistically signiifcant. By WB experiments, we found that the combination of drug treatment could inhibit Bcl-2, survivin protein expression induced by cisplat-in, while enhancing the pro-apoptosis death proteins Bax, Caspase-3 expression. Conclusion：The best choice for morphine dosage can signiifcantly enhance proliferation and apoptosis function of cisplatin on lung cancer cell. This may inhibit the proliferation of lung cancer cells by promoting key apoptotic protein Bax and Caspase-3 ex-pression pathway.