复方肠泰方通过抑制Akt/mTOR/p70S6K通路诱导CT26细胞自噬的研究  

Study on Compound Changtai Formula Inducing Autophagy in CT26 Cells through Inhibition of Akt/mTOR/p70S6K Pathway

作　　者：刘芮奇 李灵常[1,2] 崔莉[1,2] 孙娥[1,2] LIU Ruiqi;LI Lingchang;CUI Li;SUN E(The Third Clinical Medical College,Nanjing University of Chinese Medicine,Nanjing 210028,China;Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing 210028,China)

机构地区：[1]南京中医药大学第三临床医学院,江苏南京210028 [2]江苏省中医药研究院,江苏南京210028

出　　处：《中医药信息》2022年第10期1-6,共6页Information on Traditional Chinese Medicine

基　　金：国家中医临床研究基地开放课题一般项目(JD2019SZXYB04)。

摘　　要：目的:研究复方肠泰方对结肠癌CT26细胞增殖和自噬的影响及其可能的作用机制。方法:采用MTT法检测不同浓度复方肠泰方对CT26细胞增殖的影响,MDC染色法和透射电镜观察复方肠泰方干预CT26细胞后是否形成自噬体。蛋白质印迹法检测自噬相关蛋白LC3和Beclin-1的蛋白表达。采用自噬诱导剂雷帕霉素研究复方肠泰方是否诱导CT26细胞自噬性死亡。采用蛋白质印迹法检测Akt/mTOR/p70S6K通路相关蛋白表达水平。结果:复方肠泰方可以抑制CT26细胞的增殖;透射电镜观察到复方肠泰方干预的细胞中有类似自噬溶酶体结构;MDC结果显示,与空白对照组相比,复方肠泰方组细胞荧光强度增强;复方肠泰方能增加LC3Ⅱ/LC3Ⅰ和Beclin-1的表达水平(P<0.05);与复方肠泰方组相比,复方肠泰方和雷帕霉素共同处理使CT26细胞的病死率显著增加(P<0.05);此外,复方肠泰方干预后,CT26细胞Akt、mTOR和p70S6K的蛋白表达基本不变,p-Akt、p-mTOR和p-p70S6K的蛋白表达较空白对照组显著降低(P<0.05)。结论:复方肠泰方可能通过抑制Akt/mTOR/p70S6K通路诱导结肠癌CT26细胞自噬,从而发挥抗肿瘤作用。Objective:To explore the effect of Compound Changtai Formula on proliferation and autophagy of colon cancer CT26 cells and its possible mechanism.Methods:MTT was used to detect the effects of Compound Changtai Formula on the proliferation of CT26 cells.MDC staining and transmission electron microscope(TEM)were used to observe whether the autophagosomes were formed in CT26 cells after the intervention of Compound Changtai Formula,which would be stained by MDC staining.Western blot was used to detect the expressions of autophagy related proteins of LC3 and Beclin-1.The autophagy inducer Rapamycin was used to explore whether Compound Changtai Formula could induce autophagic death in CT26 cells.The protein levels related to Akt/mTOR/p70S6K pathway were detected by Western blot.Results:Compound Changtai Formula could inhibit the proliferation of CT26 cells.TEM observed the structure of autophagolysosome in the cells intervened by Compound Changtai Formula.MDC results showed that Compound Changtai group enhanced the fluorescence intensity of the cells,when compared with the blank control group.Compound Changtai Formula could up-regulate LC3Ⅱ/LC3Ⅰratio and Beclin-1 level(P<0.05).Compared with Compound Changtai Formula,the combined intervention of Compound Changtai Formula and Rapamycin significantly increased the mortality of CT26 cells(P<0.05).In addition,after the intervention of Compound Changtai Formula,the levels of Akt,mTOR and p70S6K were basically unchanged in CT26 cells,whereas the protein expressions of p-Akt,p-mTOR and p-p70S6K were significantly decreased compared with those in the blank control group(P<0.05).Conclusion:Compound Changtai Formula may induce autophagy of CT26 cells by inhibiting the Akt/mTOR/p70S6K pathway,thus producing anti-tumor effect.

关 键 词：复方肠泰方 细胞自噬 Akt/mTOR/p70S6K通路 结肠癌 CT26细胞 

分 类 号：R285[医药卫生—中药学]