扶正抗癌方通过PTEN/PI3K/Bad通路调控肺癌A549细胞增殖与凋亡

目的:观察扶正抗癌(FZKA)方对肺癌细胞A549增殖的调控作用,探讨其作用的分子机制。方法:以非小细胞肺癌细胞A549为研究对象,以FAKA方0.8,1.2,1.6,2.0,2.4,2.8,3.2 g·L~(-1)干预,空白组,分别孵育24,48,72 h后,设立采用噻唑蓝(MTT)比色法检测FZKA方对A549细胞活力的影响;以FAKA方1.2,1.6,2.0 g·L~(-1)干预,设立空白组,培养24 h后,采用实时荧光定量聚合酶链式反应(Real-time PCR)检测FZKA方对A549细胞人第10号染色体同源丢失性磷酸酶-张力蛋白(phosphatase and tensin homolog deleted on chromosome ten,PTEN)mRNA水平的影响;孵育24 h后,采用蛋白质免疫印迹法(Western blot)分析FZKA方对PTEN,磷脂酰肌醇3-激酶(phosphatidylinositol-3-kinase,PI3K),磷酸化的促凋亡蛋白Bad(Phospho-Bad,p-Bad)表达的影响并探讨其相关性。结果:与空白组比较,FZKA方能明显抑制A549细胞增殖,处理24 h后,FZKA从0.8 g·L~(-1)开始,处理48,72 h后,FZKA方从0.4 g·L~(-1)开始,细胞存活率明显降低(P0.05,P0.01);与空白组比较,处理24 h后,FZKA方从0.8 g·L~(-1)开始以浓度依赖性上调PTEN mRNA和蛋白的表达(P0.05,P0.01),FZKA方从1.6 g·L~(-1)开始以浓度依赖性下调PI3K蛋白表达(P0.05,P0.01),从2 h开始以时间依赖性上调p-Bad蛋白的表达(P0.05,P0.01)。结论:FZKA方可通过上调PTEN来调控下游PI3K/Akt/Bad通路相关蛋白的表达进而抑制A549的增殖,促进肺癌细胞凋亡。

Effect of FZKA Decoction in Regulating A549 Proliferation and Apoptosis Via PTEN/PI3K/Bad

Objective: To observe the effect of Fuzheng Kangai (FZKA) decoction on the proliferation and apoptosis, in order to discuss the molecular mechanism of apoptosis induced by FZKA decoction in A549 cells. Method: Non-small lung cancer cells (A549 cells) were taken as the research objects and treated for 24, 48, 72 h with 0 (blank group), 0.8, 1.2, 1.6, 2.0, 2.4, 2.8, 3.2 g·L^-1 FAKA decoction. Then, methyl-thiazolyl-tetrazolium (MTT) assay was used to detect the effect of FZKA decoction on the cell proliferation and apoptosis. A549 cells were treated for 24 h with 0 (blank group), 1.2, 1.6, 2.0 g·L^-1. Then, quantitative Real-time PCR was used to detect the effect of FZKA decoction on PTEN mRNA expression. A549 cells were treated for 24 h with 0 (blank group), 0.8, 1.2, 1.6, 2.0 g·L^-1. Then, Western blot assay was used to detect the expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN), phosphatidylinositol-3-kinase (PI3K) and phospho-Bad (p-Bad), and discuss the relationship between them. Result: Compared with blank group, the proliferation of A549 cells were inhibited by FZKA decoction in concentration-dependent and time-dependent manners (P < 0.01). Compared with blank group, mRNA and protein expressions of PTEN were up-regulated in a concentration-dependent manner (P < 0.05, P < 0.01). Compared with blank group, the protein expression of PI3K was down-regulated in a concentration-dependent manner (P < 0.05, P < 0.01), and p-Bad was down-regulated in a time-dependent manner (P < 0.05, P < 0.01). Conclusion: Apoptosis is induced by FZKA decoction by up-regulating PTEN and regulating PI3K/Akt/Bad pass way in A549 cells.