雌激素通过GPR30-AMPK-mTOR通路诱导Ishikawa细胞自噬增强细胞活力（英文）  

Estrogen Enhances Ishikawa Cell Viability by Inducing Autophagy via the GPR30-AMPK-mTOR Pathway

作　　者：何磊[1] 胡惠军[2] 潘勇权 谭秋芬[2] 林小龙[2] 

机构地区：[1]乐山市市中区人民医院病理科,四川乐山614000 [2]广州医科大学附属惠州医院(惠州市第三人民医院)病理科,广东惠州516002

出　　处：《中国生物化学与分子生物学报》2017年第4期353-361,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：Supported by National Natural Science Foundation of China(No.81600342);Science and Technology Program of Guangdong(No.A2014810;No.A2015620)~~

摘　　要：雌激素是子宫内膜癌发生发展的重要诱导因子,但关于其在子宫内膜癌中的作用机制目前仍不明确。自噬对细胞的存活具有重要的调节作用,研究发现其在子宫内膜癌发生发展的过程中起重要的调节作用。本文通过探讨雌激素对子宫内膜癌细胞自噬的影响,深入地了解雌激素促进子宫内膜发展的机制,并明确GPR30-AMPK-mT OR通路在其中的作用。MTT及透视电镜的结果显示,雌激素可以诱导细胞的自噬及增强细胞的活力,而这种作用具有一定的时间及浓度依赖性。同时,蛋白质印迹及实时定量PCR结果显示雌激素可以促进LC3、p-AMPK的表达,并且抑制P62、pmT OR的表达,表明雌激素可以激活AMPK/mT OR通路。沉默G蛋白偶联受体30(GPR30)后,结果显示雌激素诱导细胞的自噬及细胞活力的作用被逆转,并且可以抑制AMPK/mT OR通路的激活,而G-1结果与之相反,表明雌激素通过GPR30激活AMPK/mT OR通路,诱导自噬及细胞活力。此外,加入AMPK抑制剂compound C,可以抑制雌激素诱导细胞的自噬及细胞活力的能力,并且促进P62、p-mT OR表达,降低LC3及p-AMPK表达,表明雌激素通过激活AMPK/mT OR激活细胞自噬及增强细胞活力。同时细胞预先加入自噬抑制剂3-MA或转染ATG5siRNA,可以降低雌激素增强细胞的活力,表明雌激素通过诱导自噬增强细胞活力。综合以上结果,雌激素通过GPR30-AMPK-mT OR通路诱导细胞的自噬增强细胞的活力。Although estrogen is a key inducer of endometrial carcinoma, its mechanism of action remains unclear. Autophagy plays a role in the occurrence and development of endometrial carcinoma. In the present study, we combined these elements and investigated whether estrogen induced cellular autophagy of endometrial carcinoma cells through the GPR30-AMPK-mTOR pathway. The MTT method and transmission electron microscopy （TEM） revealed that estrogen induced autophagy and increased cellular activity of Ishikawa cells in a dose- and time-dependent manner. Meanwhile, Western blot and real-time PCR results showed that estrogen increasing the expression of p-AMPK, LC3 and inhibited p-roTOR and P62 expression. These results indicated that estrogen could activate AMPK/mTOR signal. After GPR30 silence, the results showed that estrogen-induced autophagy and enhancement of cell viability were reversed, and the activation of AMPK/mTOR pathway was inhibited, whereas the G-1 results were the opposite. These findings indicated that estrogen-induced autophagy and cell viability by activating AMPK/mTOR pathway through GPR30. In addition, adding AMPK inhibitor compound C inhibited estrogen-induced cell autophagy and cell viability, and promoted P62 and p-roTOR expression, but reduced the levels of LC3 and p-AMPK. These results suggested that estrogen activated AMPK/mTOR induced autophagy and enhanced cell viability. At the same time, addition of 3-MA or transfection of ATG5 siRNA could reduce estrogen-enhanced cellular activity, indicating that estrogen enhanced cell viability by inducing autophagy. Based on the above results, estrogen enhances Ishikawa cell viability by inducing autophagy via the GPR30-AMPK-mTOR pathway.

关 键 词：自噬 子宫内膜癌 G蛋白偶联受体30 

分 类 号：R362[医药卫生—病理学] R711[医药卫生—基础医学]