马钱子碱通过调控PI3K/Akt/mTOR通路对肺腺癌细胞的抑制作用研究

Inhibitory effect of brucine on lung adenocarcinoma cells by regulating the phosphoinositide-3 kinase/protein kinase-B/mammalian target of rapamycin pathway

目的:探究马钱子碱对非小细胞肺癌的影响及其作用机制。方法:采用CCK-8法和划痕试验法研究马钱子碱抑制A549细胞的增殖、迁移能力,Western blot法检测各组细胞磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/Akt/mTOR)通路相关的蛋白表达,ELISA法检测白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)的含量。结果:CCK-8试验结果显示,给药组对A549细胞增殖有明显抑制作用;划痕试验显示,与空白组比较,给药组细胞迁移率明显下降(P<0.01);Western blot结果显示,给药干预后可显著下调p-Akt、p-mTOR的表达(P<0.001),同时给药组的mTOR、Akt、PI3K蛋白表达下调(P<0.05);ELISA试验结果显示,采用马钱子碱进行干预后,细胞上清IL-6和TNF-α含量均显著升高(P<0.01或P<0.001)。结论:马钱子碱可有效抑制非小细胞肺癌细胞的增殖、迁移,可能是通过抑制PI3K/Akt/mTOR通路而起作用,并通过调节TNF-α含量来达到抗肿瘤的作用。

Objective: To investigate the effect of brucine on non-small cell lung cancer and its mechanism of action.Methods: CCK-8 assay and scratch test were used to investigate the effect of brucine in inhibiting the proliferation and migration abilities of A549 cells;Western blot was used to measure the expression of proteins associated with the phosphoinositide-3 kinase/protein kinase-B/mammalian target of rapamycin(PI3K/Akt/mTOR) pathway;ELISA was used to measure the content of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α).Results: CCK-8 assay showed that the drug administration group showed a significant inhibitory effect on the proliferation of A549 cells, and the scratch test showed that compared with the blank group, the drug administration group had a significant reduction in cell migration rate(P<0.01).Western blot showed that drug intervention significantly downregulated the protein expression of p-Akt and p-mTOR(P<0.001),and the drug administration group had significantly downregulated protein expression of mTOR,Akt, and PI3K(P<0.05).ELISA showed that there were significant increases in the content of IL-6 and TNF-α in supernatant after intervention with brucine(P<0.01 or P<0.001).Conclusion: Brucine can effectively inhibit the proliferation and migration of non-small cell lung cancer cells, possibly by inhibiting the PI3K/Akt/mTOR pathway and regulating the content of TNF-α to exert an antitumor effect.