芪杉胶囊对肺癌A549细胞PI3KAKt信号通路影响的研究

Effect of Qishan Capsule on PI3KAKt Signaling Pathway in Lung Cancer A549 Cells

目的:观察芪杉胶囊对肺癌A549细胞PI3KAKt信号通路的影响。方法:分别运用MTT法检测含药血清对A549细胞增殖的影响;Annexin V-PE/7-AAD双染法检测含药血清对A549细胞凋亡的影响;Western-blot法检测含药血清对A549细胞凋亡相关蛋白表达的影响;RT-PCR法检测含药血清对A549细胞凋亡相关基因表达的影响。结果:芪杉胶囊含药血清高、中、低剂量组分别作用于A549细胞时,均表现为抑制细胞增殖作用,其抑制率分别为24.26%,16.44%和10.89%,含药血清浓度越大,对细胞的抑制作用越显著。与空白组相比,芪杉胶囊含药血清高、中、低剂量组及顺铂组细胞凋亡率显著升高,且含药血清浓度越大,对细胞凋亡的作用越显著。芪杉胶囊含药血清高、中、低剂量组及顺铂组Caspase-3、Caspase-9、Cytochrome c、AKT、m TOR及PI3K蛋白和基因相对表达量与空白组相比,表达量降低,且含药血清浓度越大,对A549细胞凋亡相关蛋白和基因表达的影响越显著。结论:芪杉胶囊通过影响A549细胞凋亡相关Caspase-3、Caspase-9、Cytochrome c、AKT、m TOR及PI3K蛋白和基因表达,从而抑制细胞增值、促进细胞凋亡。

Objective： To observe the effect of Qishan Capsule on PI3KAKt signaling pathway in lung cancer A549 cells.Methods： The proliferation and apoptosis of A549 cells intervend with medicated serum were detected by MTT method and Annexin V-PE/7-AAD double staining method respectively. Protein and gene expressions related to apoptosis of A549 cells were tested by Western-blot assay and RT-PCR assay. Results： It is found out that different concentration of serum contaning Qishan Capsule had inhibiting effect on the proliferation of A549 cells; the inhibition rate was 24. 26%,16. 44% and 10. 89%,the higher concentration of the serum was,the more significant inhibitory effect it had on the proliferation of A549 cells. In terms of apoptosis rate,it was significantly increased in serum-Qishan Capsule groups and in cisplatin group,compared to that in the blank control group; the higher concentration of the serum was,the more significant effect it had on apoptosis. In terms of expressions of Caspase-3,Caspase-9,Cytochrome C,AKT,m TOR and PI3K,they were significantly decreased in serum-Qishan Capsule groups and in cisplatin group,compared to those in the blank control group; the higher concentration of the serum was,the more significant decrease was seen on the expressions. Conclusion： Qishan Capsule can inhibit the proliferation and promote apoptosis of A549 cells by regulating the expressions of Caspase-3,Caspase-9,Cytochrome C,AKT,mTOR and PI3 K.