苦参碱通过抑制PI3K/Akt/mTOR信号通路诱导食管癌Ec109细胞自噬

Matrine induces autophagy in Ec109 cells via inhibiting PI3K/Akt/mTOR signaling pathway

目的探究苦参碱对食管癌Ec109细胞的自噬作用及其机制。方法体外培养食管癌Ec109细胞,分别经不同浓度苦参碱处理后,倒置相差显微镜下观察细胞形态学变化;四甲基偶氮唑蓝法检测细胞增殖抑制率;Western blotting检测自噬相关蛋白LC3-Ⅱ及PI3K/Akt/mTOR信号通路相关蛋白p-mTOR、p-Akt和p-p70S6K的表达;实时荧光定量PCR检测自噬相关基因Beclin1 mRNA的表达。结果倒置显微镜下可见,食管癌Ec109细胞经苦参碱处理后胞体缩小,胞内可发现不均一的细颗粒状物质。不同浓度的苦参碱处理48 h,细胞增殖抑制率随着苦参碱浓度的增加逐渐升高(P<0.05)。Western blotting检测结果显示,LC3-Ⅱ的表达随苦参碱浓度的升高而增高,但p-mTOR、p-Akt和p-p70S6K的表达均随苦参碱浓度的增高而降低;实时荧光定量PCR结果显示,苦参碱处理使自噬基因Beclin1 mRNA的表达上调。结论苦参碱诱导食管癌细胞自噬可能是通过PI3K/Akt/mTOR信号通路介导的。

Objective To investigate the autophagic effect of matrine on Ec109 cells and its possible mechanism.Methods Ec109 cells were cultured in vitro and their morphological changes were observed by inverted phase contrast microscopy after they were treated with matrine at different concentrations.The inhibition rate was detected by methyl thiazolyl tetrazolium.Western blotting was used to detect the expressions of LC3-Ⅱ,p-mTOR,p-Akt and p-p70S6K.RT-qPCR was used to detect the expression of Beclin1.Results Under the inverted phase contrast microscope,the volume of Ec109 cells decreased after matrine treatment,and non-homogeneous granular substances could be found in the cells.Western blotting results showed that the expression of LC3-Ⅱincreased while the expressions of p-mTOR,p-Akt and p-p70S6K decreased with the increase of matrine concentration(P<0.05).RT-qPCR showed that the expression of Beclin1 was up-regulated by matrine.Conclusion Matrine can induce autophagy in Ec109 cells and this process may be mediated by PI3K/Akt/mTOR signaling pathway.