化疗介导mTOR/AKT信号通路调控炎症因子分泌抑制非小细胞肺癌发展

Chemotherapy-mediated mTOR/AKT signaling pathway regulates the secretion of inflammatory cytokines to inhibit the development of NSCLC

目的探讨化疗介导mTOR/AKT信号通路调控炎症因子分泌抑制非小细胞肺癌发展及相关机制。方法以化疗药物紫杉醇(Paclitaxel,PAC)梯度浓度(0、5、10、50μmol)处理非小细胞肺癌细胞系A549细胞,Western blot检测p-mTOR和pAKT表达变化。进一步使用mTOR/AKT信号通路激动剂SC79进行功能逆转实验,荧光定量PCR检测M1型炎症因子IL-6和iNOS以及M2型炎症因子Arg-1和CD206表达变化;CCK-8和Edu染色检测细胞增殖情况,Tunel染色检测细胞凋亡情况,Western blot检测凋亡相关蛋白Bax/Bcl2和Cleaved-caspase3/Caspase3表达变化。结果化疗药物紫杉醇可抑制p-mTOR和pAKT表达,并且具有明显的梯度相关性(P<0.05)。紫杉醇可明显逆转SC79诱导的A549细胞M1型炎症因子IL-6和iNOS表达的升高(P<0.05),M2型炎症因子Arg-1和CD206表达的降低(P<0.05)。此外,紫杉醇可明显抑制SC79诱导的A549细胞增殖增多(P<0.05)、凋亡减少(P<0.05)及Bax/Bcl2和Cleaved-caspase3/Caspase3表达降低(P<0.05)。结论化疗药物紫杉醇通过mTOR/AKT信号通路调控炎症因子分泌,达到抑制非小细胞肺癌发展的效果。

This study was designed to investigate the regulatory effects of chemotherapy-mediated mTOR/AKT signaling pathway on the secretion of inflammatory cytokines against the development of non-small cell lung cancer(NSCLC)and to explore the related mechanisms.NSCLC cell line A549 cells were treated with the chemotherapy drug paclitaxel(PAC)at gradient concentrations of 0,5,10 and 50μmol,and the expression of PmTOR and P-Akt were detected by Western blotting.mTOR/AKT signaling pathway agonist SC79 was used for functional reversal,and the expression changes of M1 inflammatory cytokines IL-6 and iNOS as well as M2 inflammatory cytokines Arg-1 and CD206 were detected by qPCR.The cell proliferation and apoptosis were detected by CCK-8,Edu staining and Tunel staining.Data showed that paclitaxel inhibited the expression of P-mTOR and P-AKT in a dose dependent manner(P<0.05).Fluorescence quantitative PCR showed that paclitaxel significantly reversed SC79-induced expression of IL-6 and iNOS as well as SC79-suppressed expression of Arg-1 and CD206 in A549 cells(P<0.05).Furthermore,paclitaxel significantly inhibited the proliferation of SC79-treated A549 cells,promoted the apoptosis of the cells,also up-regulated the expression of Bax/Bcl2 and cleaved caspase3/caspase3 in SC79-treated A549 cells(P<0.05).These results indicated that the chemotherapeutic drug paclitaxel can inhibit the development of NSCLC by regulating the secretion of inflammatory cytokines through the mTOR/AKT signaling pathway.