莪术醇对肺癌A549细胞凋亡诱导因子、 聚ADP核糖聚合酶及Caspase-3表达的影响

目的:观察莪术醇干预后,体外培养A549细胞凋亡诱导因子(apoptosis induce factor,AIF)和聚ADP核糖聚合酶poly(ADP-ribose)polymerase,PARP]的表达情况,探讨莪术醇对非半胱天冬酶(caspase)依赖细胞凋亡的分子机制。方法:以不同质量浓度的莪术醇(12.5,25,50,100 mg.L-1)及阴性组处理肺癌细胞,药物作用48 h后提取各组蛋白;采用Westernblotting法分别检测各组AIF,PARP,caspase-3蛋白表达,并进行统计学分析。结果:经莪术醇处理后,阴性组的A549细胞的PARP相对蛋白表达量为0.545 3,AIF的相对蛋白表达量为0.358 6,而100 mg.L-1加药组的相对蛋白表达量分别为1.661 3和2.212 4,蛋白表达均随药物浓度的增加而增多,药物组与阴性组相比差异极显著(P<0.01),而caspase-3的前体(32×103)表达受药物影响较小,其激活体(17×103)基本不表达。结论:莪术醇可能主要是通过上调PARP蛋白的表达,进而将AIF转位到细胞核而引起凋亡的非依赖caspase途径来介导肺癌A549细胞的凋亡。

Effects of Curcumol on Apoptosis Induce Factor,Poly ADP-Ribose Polymerase and Caspase-3 in Lung Cancer Cell Line A549

Objective: To study the effects of curcumol on the expression of apoptosis induce factor(AIF),poly ADP-ribose polymerase(PARP)and caspase-3 protein in lung cancer cell A549. Method: A549 cells were treated with different concentrations(0,12.5, 25, 50,100 mg·L^-1)of curcumol for 48 hours, then the total protein was extracted from each group respectively. The expression of AIF,PARP and caspase-3 was detected by Western blotting. Result: The PARP and AIF protein expressions were significantly elevated in the treatment groups(1.661 3 and 2.212 4 respectively)compared with the model group(0.545 3 and 0.358 6 respectively), in a dose dependent manner(P<0.01). The caspase-3 precursor(32×10³)protein expression showed no significance difference among all the groups. Conclusion: Curcumol could induce apoptosis through caspase-independent pathway in human lung cancer cell A549, whose mechanism might be linked to up-regulation of the PARP protein and translocation of AIF.