长链非编码RNA NEAT1通过miR-34a靶向GAS1激活PI3K-AKT信号通路对宫颈癌的增殖、侵袭和上皮细胞-间充质转化的影响研究  

LncRNA NEAT1 activates PI3K-AKT signaling pathway through miR-34a targeting GAS1 to promote the proliferation, invasion and EMT of cervical cancer

作　　者：刘莹 谢锦霞 李恒[2] LIU Ying;XIE Jinxia;LI Heng(Department of Obstetrics and Gynecology,Xi′an Fourth Hospital,Xi'an 710004,Shaanxi,China;School of Basic Medical Sciences,Xi′an Jiaotong University,Xi'an 710049,Shaanxi,China)

机构地区：[1]西安市第四医院妇产科,西安710004 [2]西安交通大学基础医学院,西安710049

出　　处：《中国性科学》2021年第5期59-63,共5页Chinese Journal of Human Sexuality

基　　金：陕西省重点研发计划项目(2019SF-205)。

摘　　要：目的探讨长链非编码RNA(lncRNA)NEAT1对宫颈癌细胞增殖、侵袭和上皮细胞-间充质转化(EMT)的影响和机制研究。方法选取2019年10月购自上海生物细胞研究所的宫颈癌细胞株(Hela细胞)和人宫颈上皮永生化细胞株(H8细胞)作为研究对象。通过实时荧光定量聚合酶链反应(RT-qPCR)检测NEAT1在Hela细胞和H8细胞中的差异表达,分析NEAT1对Hela细胞增殖、侵袭和EMT的影响。miRanda和双荧光素酶报告基因实验分析NEAT1和miR-34a之间的关系,检测NEAT1通过miR-34a对Hela细胞增殖、侵袭与EMT的影响,TargetScan和双荧光素酶报告基因实验分析miR-34a和GAS1的关系,检测miR-34靶向GAS1激活PI3K-AKT信号通路对Hela细胞增殖、侵袭和EMT的影响。分别用MTT、细胞侵袭实验和Western blot检测细胞的增殖、侵袭和EMT能力。结果 Hela细胞中NEAT1的表达上调(P<0.01),miR-34a表达下调(P<0.01),敲低NEAT1抑制了Hela细胞增殖、侵袭与EMT,NEAT1 3′UTR与miR-34a特异性结合。同时敲低NEAT1和miR-34a的表达,NEAT1表达的下调能部分逆转miR-34a下调对细胞功能的影响。同时过表达NEAT1和miR-34a, NEAT1能部分逆转miR-34a上调对细胞功能的影响。miR-34a和GAS13′UTR特异性结合,敲低GAS1抑制了Hela细胞增殖、侵袭与EMT,干扰GAS1抑制了Hela细胞p-PI3K、p-AKT蛋白的表达。和干扰miR-34a相比,同时抑制GAS1和miR-34a的表达能够逆转p-PI3K、p-AKT蛋白的上升。MK2206抑制了Hela细胞增殖、侵袭与EMT,而IGF-1促进了细胞的增殖、侵袭与EMT。结论 NEAT1通过miR-34a靶向GAS1激活了PI3K-AKT信号通路,从而促进宫颈癌的发展。Objective To investigate the effect of long noncoding RNA(lncRNA) NEAT1 on proliferation, invasion and epithelial-mesenchymal transition(EMT) of cervical cells and the underlying mechanism. Methods Cervical cancer cell strain(Hela cells) and human cervical epithelial immortalized cell strain(H8 cells) purchased from Shanghai Institute of Biochemistry and Cell Biology in October 2019 were selected as research objects. RT-qPCR was used to detect the expression of NEAT1 in Hela and H8 cells. The effects of NEAT1 on the proliferation, invasion and EMT of Hela cell were analyzed. The miRanda and Dual luciferase reporter gene assay were used to detect the correlation between NEAT1 and miR-34 a. The effect of NEAT1 on the proliferation, invasion and EMT of Hela cell through miR-34 a was analyzed. The TargetScan database and Dual luciferase reporter gene were used to analyze the correlation between miR-34 a and GAS1. The effect of miR-34 targeting GAS1 and activating the PI3 K-AKT signaling pathway on the proliferation, invasion and EMT of Hela cells was analyzed. Cell proliferation, invasion and EMT were measured by MTT, cell invasion assay and Western blot, respectively. Results In Hela cells, the expression of NEAT1 was up-regulated(P<0.01), the expression of miR-34 a was down-regulated(P<0.01). The down-regulation of NEAT1 inhibited Hela cell proliferation, invasion and EMT. NEAT1 3′UTR specifically bonded to the miR-34 a. Knocking down the expression of NEAT1 and miR-34 a at the same time, the down-regulation of NEAT1 expression partially reversed the effect of miR-34 a down-regulation on cell function. Overexpression of NEAT1 and miR-34 a at the same time, the overexpression of NEAT1 expression partially reversed the effect of miR-34 a overexpression on cell function. miR-34 a specifically bonded to GAS13′UTR, the down-regulation of GAS1 inhibited Hela cell proliferation, invasion and EMT. The down-regulation of GAS1 expression inhibited the expression of p-PI3 K and p-AKT proteins in Hela cells. Compared with

关 键 词：NEAT1 MIR-34A GAS1 PI3K-AKT 宫颈癌 上皮细胞-间充质转化 

分 类 号：R711[医药卫生—妇产科学]