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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。
Estrogen is an important inducer of the development of endometrial cancer, but its mechanism of action in endometrial cancer remains unclear. Autophagy plays an important role in the regulation of cell survival. It has been found that autophagy plays an important regulatory role in the development of endometrial cancer. This article explored the effect of estrogen on autophagy in endometrial cancer cells, and further explored the mechanism by which estrogen promotes the development of endometrium and clarified the role of GPR30-AMPK-mT OR pathway in it. MTT and fluoroscopy results show that estrogen can induce cell autophagy and enhance cell viability, and this effect has a certain time and concentration-dependent. Meanwhile, western blotting and real-time quantitative PCR showed that estrogen can promote the expression of LC3 and p-AMPK, and inhibit the expression of P62 and pmT OR, indicating that estrogen can activate AMPK / mT OR pathway. Silencing G-protein coupled receptor 30 (GPR30) showed that estrogen-induced autophagy and cell viability were reversed and AMPK / mT OR signaling was inhibited, whereas G-1 showed the opposite result, This indicates that estrogen activates AMPK / mT OR through GPR30 and induces autophagy and cell viability. In addition, the addition of AMPK inhibitor compound C can inhibit the estrogen-induced cell autophagy and cell viability, and promote P62, p-mT OR expression, reduce LC3 and p-AMPK expression, indicating that estrogen through the activation of AMPK / mT OR activates cell autophagy and enhances cell viability. At the same time cells pre-joined with autophagy inhibitor 3-MA or transfected with ATG5siRNA, can reduce the estrogen-enhanced cell viability, indicating that estrogen through the induction of autophagy to enhance cell viability. Taken together, estrogen induces cell autophagy via the GPR30-AMPK-mT OR pathway to enhance cell viability.