敲低葡萄糖-6-磷酸酶催化亚基(G6PC)阻断AKT/mTOR通路抑制宫颈癌HeLa细胞增殖、侵袭和迁移  

G6PC knockdown blocks AKT/mTOR pathway to inhibit proliferation,invasion and migration of cervical cancer HeLa cells

作　　者：邓春玲 朱坤 刘涛瑞 陈丽艳 DENG Chunling;ZHU Kun;LIU Taorui;CHEN Liyan(Cancer Research Center,College of Medicine,Yanbian University;Jilin Provincial Key Laboratory of Gynecological Oncology Bioinformatics,Yanji 133002,China)

机构地区：[1]延边大学医学院肿瘤研究中心 [2]吉林省妇科肿瘤生物信息学重点实验室,吉林延吉133002

出　　处：《细胞与分子免疫学杂志》2021年第6期520-526,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81660436,81460399)。

摘　　要：目的探讨葡萄糖-6-磷酸酶催化亚基(G6PC)对宫颈癌HeLa细胞的增殖、侵袭和迁移性能的影响及可能的分子机制.方法采用RNA干扰技术(RNAi)沉默宫颈癌HeLa细胞中G6PC的表达,Western blot法检测沉默效果.敲低G6PC,噻唑蓝(MTT)法和集落形成实验检测宫颈癌HeLa细胞增殖,TranswellTM实验和划痕愈合实验检测细胞的侵袭和迁移能力,血管形成实验检测血管生成能力;Western blot法检测上皮间质转化(EMT)相关蛋白上皮钙黏蛋白(E-cadherin)、波形蛋白(vimentin)、snail、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)、血管内皮生长因子(VEGF)蛋白表达.结果RNAi可有效下调宫颈癌HeLa细胞G6PC的表达,敲低G6PC能明显抑制宫颈癌HeLa细胞的增殖、侵袭、迁移和血管生成,同时抑制EMT进程以及AKT、mTOR蛋白的磷酸化.结论敲低G6PC抑制HeLa细胞的增殖、侵袭、迁移、血管形成和EMT进程,与阻断AKT/mTOR信号通路相关.Objective To explore the effect of glucose-6-phsophatase, catalytic subunit(G6PC) on the proliferation, migration and invasion of cervical cancer HeLa cells and the possible molecular mechanism. Methods RNA interfering(RNAi) was used to knockdown the expression of G6PC in HeLa cells, and the silencing effect of protein was confirmed by Western blotting. MTT assay and plate clony formation assay were performed to detect the effect of G6PC knockdown on the proliferation of HeLa cells;scratch healing assay and TranswellTM chamber assay were applied to observe the effect of G6PC knockdown on the invasion and migration abilities of HeLa cells;the tube-formation assay was used to detect the effect of G6PC knockdown on the angiogenesis ability of HeLa cells;the expression levels of epithelial-mesenchymal transition(EMT)-related proteins and AKT/mTOR signaling pathway-related proteins were determined by Western blotting. Results The expression of G6PC was effectively silenced by RNAi technology. G6PC knockdown obviously inhibited the proliferation, migration and angiogenesis of HeLa cells. Meanwhile, G6PC knockdown suppressed the EMT process, the phosphorylation of AKT and mTOR proteins. Conclusion G6PC knockdown can effectively inhibit the proliferation, migration, angiogenesis and EMT process of HeLa cells, which is related to the blocked AKT/mTOR signaling pathway.

关 键 词：葡萄糖-6-磷酸酶催化亚基(G6PC) 宫颈癌 细胞增殖 上皮间质转化(EMT) 血管形成 蛋白激酶B(AKT) 哺乳动物雷帕霉素靶蛋白(mTOR) 

分 类 号：R737.33[医药卫生—肿瘤] R711.74[医药卫生—临床医学]