扶正抑瘤汤通过PI3K/Akt/mTOR信号通路抑制非小细胞肺癌增殖、凋亡及自噬

目的 探讨扶正抑瘤汤对非小细胞肺癌细胞增殖、侵袭、迁移、凋亡、自噬以及PI3K/Akt/mTOR信号通路的影响。方法 采用不同浓度（12.5 mg/mL、25 mg/mL、50 mg/mL、100 mg/mL、200 mg/mL、400 mg/mL）扶正抑瘤汤体外培养非小细胞肺癌A549细胞， CCK-8法测定细胞增殖抑制率；Transwell小室侵袭实验检测细胞侵袭能力；划痕实验检测细胞迁移能力；流式细胞仪检测细胞凋亡率；Western blot检测细胞自噬相关蛋白以及PI3K/Akt/mTOR信号通路相关蛋白的表达水平；透射电镜下观察自噬溶酶体形成情况。结果 200 mg/mL和400 mg/mL扶正抑瘤汤作用后A549细胞增殖抑制率增加（均P<0.05）。200 mg/mL扶正抑瘤汤作用后A549细胞侵袭和迁移能力降低（均P<0.05）；细胞凋亡率增加（P<0.05）；Beclin-1、LC3Ⅱ/Ⅰ蛋白的表达水平升高（均P<0.05），P62蛋白的表达水平降低（P<0.05）；A549细胞自噬溶酶体数量增加；p-PI3K/PI3K、p-Akt/Akt和p-mTOR/mTOR蛋白的表达降低（均P<0.05）。结论 扶正抑瘤汤可以抑制A549细胞增殖，并促进细胞凋亡和自噬，可能与PI3K/Akt/mTOR信号通路失活有关，是治疗非小细胞肺癌的潜在药物。

Fuzheng Yiliu decoction inhibits the proliferation,apoptosis and autophagy of non-small cell lung cancer through PI3K/Akt/mTOR signaling pathway

ObjectiveTo investigate the effects of Fuzheng Yiliu decoction on the proliferation,invasion,migration,apoptosis,autophagy and PI3 K/Akt/mTOR signaling pathway of non-small cell lung cancer.MethodsThe non-small cell lung cancer A549 cells were cultured in vitro with different concentrations(12.5 mg/mL,25 mg/mL,50 mg/mL,100 mg/mL,200 mg/mL,400 mg/mL)of Fuzheng Yiliu decoction,and the proliferation inhibition rate was determined by the CCK-8 assay;the invasion of cells was detected by Transwell assay;the cell migration was measured by the scratch test;the apoptosis rate was detected by the flow cytometry;the expression levels of autophagy-related proteins and PI3 K/Akt/mTOR signaling pathway-related proteins were detected by the Western blot,and the formation of autophagy lysosome was observed under the transmission electron microscope.ResultsAfter treated with 200 mg/mL and 400 mg/mL of Fuzheng Yiliu decoction,the proliferation inhibition rate of A549 cells was increased(all P<0.05).After treated with200 mg/mL of Fuzheng Yiliu decoction,the invasion and migration of A549 cells were decreased(all P<0.05);the apoptosis rate of A549 cells was increased(P<0.05);the expression levels of Beclin-1 and LC3Ⅱ/Ⅰproteins were increased(all P<0.05),while the expression levels of P62 protein was decreased(P<0.05);the number of autophagy lysosomes increased in A549 cells;the expression levels of p-PI3 K/PI3 K,p-Akt/Akt,and p-MTOR/mTOR proteins were decreased(all P<0.05).ConclusionsFuzheng Yiliu decoction can inhibit the proliferation of A549 cells and promote cell apoptosis and autophagy,which may be related to the inactivation of PI3 K/Akt/mTOR signaling pathway,and may be a potential drug for the treatment of non-small cell lung cancer.