外源性PUMA对非小细胞肺癌A549细胞增殖和凋亡的影响

Effects of Exogenous PUMA on Proliferation and Apoptosis of Non-Small Cell Lung Cancer A549 Cells

目的探讨外源性p53上调凋亡调制因子(PUMA)对非小细胞肺癌A549细胞增殖、凋亡的影响。方法将肺癌A549细胞分为Control组、空载体组、PUMA组(转染pCEP4-(HA)2-PUMA质粒),DDP组(10μg·mL-1)和PUMA+DDP组(转染pCEP4-(HA)2-PUMA质粒,加10μg·mL-1顺铂)5个组。细胞生存率和凋亡率分别用MTT法和流式细胞仪检测,细胞PUMA、Bax及Bcl-2蛋白表达水平采用Western blot方法检测。结果与Control组比较:PUMA组、DDP组及PUMA+DDP组A549细胞增殖均显著降低(P<0.01),且PUMA+DDP组降低程度最强(P<0.01);凋亡率均显著升高(P<0.01);Bax蛋白水平呈显著性升高(P<0.01),Bcl-2蛋白呈显著性下降(P<0.01)。结论外源性PUMA可抑制肺癌A549细胞增殖,促进凋亡,其机制与增加细胞Bax蛋白表达,降低Bcl-2蛋白表达有关。

ABSTRACT：Objective To investigate the effects of exogenous p53 up-regulated modulator of apoptosis（PUMA）on the proliferation and apoptosis of non-small cell lung cancer A549 cells. Methods Non-small cell lung cancer A549 cells were divided into 5 groups：control group,empty vector group,PUMA group（cells were transfected with pCEP4-（HA）2-PUMA plasmids）,DDP group（cells were treated with 10 μg＆#183;mL-1 cisplatin）,and PUMA＋DDPgroup（cells were trans-fected with pCEP4-（HA）2-PUMA plasmids and treated with 10 μg＆#183;mL-1 cisplatin）.Cell growth and apoptosis were determined by MTT assay and flow cytometry,respectively.The expression of PUMA,Bax and Bcl-2 proteins were detected by Western blot.Results Compared with control group,the proliferation of A549 cells was decreased and the apoptosis of A549 cells was increased in PUMA,DDP and PUMA＋DDP groups（P 〈0.01）.Maximum decrease in cell proliferation was observed in PUMA＋DDP group（P 〈0.01）.In addition,compared with control group,Bax pro-tein expression was up-regulated and Bcl-2 protein expression was down-regulated in PUMA, DDP and PUMA＋DDP groups（P 〈0.01）.Conclusion Exogenous PUMA can inhibit prolifera-tion and promote apoptosis through increasing Bax expression and decreasing Bcl-2 expression in non-small cell lung cancer A549 cells.