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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(P<0.05,P<0.01);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(P<0.05,P<0.01),FZKA方从1.6 g·L~(-1)开始以浓度依赖性下调PI3K蛋白表达(P<0.05,P<0.01),从2 h开始以时间依赖性上调p-Bad蛋白的表达(P<0.05,P<0.01)。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。
Objective: To observe the regulatory effect of FZKA on the proliferation of lung cancer A549 cells and to explore its molecular mechanism. Methods: The non-small cell lung cancer A549 cells were treated with FAKA 0.8, 1.2, 1.6, 2.0, 2.4, 2.8 and 3.2 g · L -1, respectively. The blank groups were incubated with 24, 48 and 72 h The effect of FZKA on the viability of A549 cells was determined by MTT colorimetric assay. After the cells were treated with 1.2, 1.6, 2.0 g · L -1 of FAKA for 24 h, The effect of FZKA on the mRNA level of PTEN in chromosome 10 of human A549 cells was detected by real-time PCR After incubation for 24 h, the effect of FZKA on PTEN, phosphatidylinositol-3-kinase (PI3K), phosphorylated pro-apoptotic protein Bad (Phospho-Bad , p-Bad) expression and explore its relevance. Results: Compared with the blank group, FZKA could significantly inhibit the proliferation of A549 cells. After treatment for 24 h, FZKA started from 0.8 g · L -1, and treated with FZKA from 0.4 g · L -1 -1), the cell viability decreased significantly (P <0.05, P <0.01). Compared with the blank group, FZKA treatment increased the expression of PTEN mRNA from 0.8 g · L -1 in 24 h (P <0.05, P <0.01). FZKA down-regulated PI3K protein expression from 1.6 g · L -1 in a concentration-dependent manner (P <0.05, P <0.01) The time-dependent up-regulation of p-Bad protein expression (P <0.05, P <0.01). Conclusion: FZKA can up-regulate PTEN to regulate the expression of PI3K / Akt / Bad signaling pathway, which can inhibit the proliferation of A549 cells and promote the apoptosis of lung cancer cells.