基于PTEN-PI3K-AKT信号通路探讨竹节参总皂苷抑制人肺腺癌A549细胞增殖和转移的机制

目的:竹节参是人参属植物,和人参成分相似,前期研究其对肺癌具有一定的抑制作用,但作用机制不清,因此,本项目拟研究竹节参皂苷对人肺癌细胞系A549增殖、迁移和侵袭能力以及PTEN-PI3K-AKT信号通路的影响。方法:CCK8法测定不同浓度和不同作用时间的竹节参皂苷对A549存活率的影响,划痕实验测定细胞迁移能力,Transwell小室测定细胞的侵袭能力,ELISA试剂盒测定培养基上清中MMP-2和MMP-9水平的变化。Western blot测定PTEN、P-PI3K和P-Akt表达的变化。结果:竹节参皂苷对A549细胞增殖具有明显的抑制作用,呈浓度和时间依赖关系,与对照组比较具有统计学差异。同时,竹节参皂苷可以浓度依赖性的抑制细胞侵袭和转移,以及MMP-2和MMP-9细胞因子的分泌。Western blot结果表明竹节参皂苷可促进PTEN蛋白表达,抑制P-PI3K和P-Akt蛋白表达,采用PTEN的特异性抑制剂SF1670证实竹节参皂苷通过抑制PTEN发挥作用。结论:竹节参皂苷可抑制肺癌A549细胞增殖、迁移和侵袭,以及分泌蛋白MMP-2和MMP-9表达,其作用机制可能是通过调控PTEN抑制PI3K和Akt磷酸化,从而发挥抗癌作用。

Study on the Mechanism of Saponins from Panaxjaponicus on Inhition of A549 Cell Proliferation and Migration through Regulating PTEN/PI3K/Akt Pathway

ABSTRACT Objective: Panax japonicus is a kind of ginseng plant and the chemical components are similar to ginseng. It has been shown that it has certain inhibitory effect on lung cancer, but the mechanism is unclear. Thus, this study was designed to study the effects of saponins from Panax japonicus (SPJ) on proliferation, migration, invasion of human lung cancer cell line A549 and PTEN-PI3K-AKT signaling pathway. Methods: The effects of SPJ at different concentration and time on the survival rate of A549 were determined by CCK8 assay. A549 cells migration ability was detected by scratch assay. The invasion ability of A549 cells was measured by Transwell chamber. The levels of MMP-2 and MMP-9 in the medium supernatant were determined by ELISA kit. The expression of PTEN, P-PI3K and P-Akt was measured by Western blot. Results: SPJ significantly inhibited the proliferation of A549 cells in a concentration- and time-dependent relationship. At the same time, SPJ inhibited the invasion, invasion and the secretion of MMP-2 and MMP-9 cytokines in A549 cells in a concentration-dependent manner. Western blot results showed that saponins promoted the expression of PTEN protein and inhibited the expression of P-PI3K and P-Akt proteins. SF1670, a specific inhibitor of PTEN, was used to confirm the effect of SPJ on inhibition of PTEN. Conclusion: SPJ inhibited the proliferation, migration and invasion of A549 cells, and the secretory protein expression of MMP-2 and MMP-9, which mechanism might through regulating PTEN and inhibiting the phosphorylation of PI3K and Akt, so as to play an anti-cancer role.