GNAS-AS1靶向miR-2116-3p基因表达调控卵巢癌细胞增殖、迁移和侵袭的分子机制研究  

The molecular mechanism of GNAS-AS1 regulating proliferation,migration and invasion of ovarian cancer cell by targeting miR-2116-3p gene expression

作　　者：宋芷霜[1] 郭宝芝[1] 刘爱珍[1] 丁俊珊 王宝金[2] Song Zhishuang;Guo Baozhi;Liu Aizhen(Department of Gynecology,Zhengzhou Women and Children's Hospital,Zhengzhou 450000)

机构地区：[1]郑州市妇幼保健院妇科,郑州450000 [2]郑州大学第三附属医院妇科河南省卵巢恶性肿瘤国际联合实验室,郑州450000

出　　处：《现代妇产科进展》2021年第4期241-246,共6页Progress in Obstetrics and Gynecology

基　　金：2019年河南省科技计划项目(No:192102310067);2019年度河南省重点研究与推广专项(No:2019-156)。

摘　　要：目的:探讨GNAS-AS1对卵巢癌细胞增殖、迁移和侵袭的影响及可能机制。方法:RT-qPCR法检测正常人卵巢上皮细胞HUM-CELL-0088及卵巢癌细胞系SKOV3、OVCAR3和A2780细胞中GNAS-AS1和miR-2116-3p表达水平。以SKOV3细胞为研究对象,分别转染GNAS-AS1小干扰RNA、miR-2116-3p模拟物或抑制剂,CCK-8法、Transwell和Western blot法分别检测下调GNAS-AS1、上调miR-2116-3p或下调miR-2116-3p对细胞增殖、迁移和侵袭及CyclinD1、MMP-2、MMP-9、N-Cadherin、Vimentin和E-Cadherin蛋白表达的影响。双荧光素酶报告基因实验验证GNAS-AS1与miR-2116-3p调控关系。结果:与HUM-CELL-0088细胞比较,卵巢癌细胞系SKOV3、OVCAR3和A2780中GNAS-AS1表达升高(P<0.05),miR-2116-3p表达降低(P<0.05)。下调GNAS-AS1表达或上调miR-2116-3p表达后,SKOV3细胞增殖、迁移和侵袭细胞数及细胞中CyclinD1、MMP-2、MMP-9、N-Cadherin和Vimentin蛋白表达水平降低(P<0.05),E-Cadherin蛋白表达水平升高(P<0.05)。GNAS-AS1在SKOV3细胞中负调控miR-2116-3p表达。下调miR-2116-3p表达或同时下调GNAS-AS1和miR-2116-3p表达后,SKOV3细胞增殖、迁移和侵袭细胞数及细胞中CyclinD1、MMP-2、MMP-9和N-Cadherin和Vimentin蛋白表达水平升高(P<0.05),E-Cadherin蛋白表达水平降低(P<0.05)。结论:GNAS-AS1在卵巢癌细胞系中表达升高,下调其表达可降低卵巢癌细胞的增殖、迁移和侵袭能力,其可能通过负调控miR-2116-3p表达发挥作用。Objective:To investigate the effect of GNAS-AS1 on the proliferation,migration and invasion of ovarian cancer cell and its possible mechanism.Methods:RT-qPCR was used to detect the expression levels of GNAS-AS1 and miR-2116-3p in normal human ovarian epithelial cells HUM-CELL-0088 and ovarian cancer cell lines SKOV3,OVCAR3 and A2780.Taking SKOV3 cells as the research objects,after GNAS-AS1 small interfering RNA,miR-2116-3p mimics or inhibitors were transfected into SKOV3 cells.CCK-8,Transwell and Western blot were used to detect the effect of down-regulating GNAS-AS1,up-regulating miR-2116-3p or down-regulating miR-2116-3p on cell proliferation,migration and invasion,and the protein expressions of CyclinD1,MMP-2,MMP-9,N-Cadherin,Vimentin and E-cadherin.The dual luciferase reporter gene experiment verified the regulatory relationship between GNAS-AS1 and miR-2116-3p.Results:Compared with HUM-CELL-0088 cells,the expressions of GNAS-AS1 in ovarian cancer cell lines SKOV3,OVCAR3 and A2780 increased(P<0.05),while the expressions of miR-2116-3p decreased(P<0.05).After down-regulating GNAS-AS1 or up-regulating miR-2116-3p,the proliferation,the number of migration and invasion of SKOV3 cells and the protein expression levels of CyclinD1,MMP-2,MMP-9,N-Cadherin and Vimentin in the cells decreased(P<0.05),while the protein expression level of E-Cadherin increased(P<0.05).GNAS-AS1 negatively regulated the expression of miR-2116-3p in SKOV3 cells.After down-regulating miR-2116-3p or simultaneously down-regulating GNAS-AS1 and miR-2116-3p,the proliferation,the number of migration and invasion of SKOV3 cells and the protein expression levels of CyclinD1,MMP-2,MMP-9,N-Cadherin and Vimentin increased in the cells(P<0.05),while the protein expression level of E-Cadherin decreased(P<0.05).Conclusion:The expression of GNAS-AS1 was increased in ovarian cancer cell lines,and down-regulating its expression could reduce the proliferation,migration and invasion of ovarian cancer cells,which may play a role by negatively regulating the ex

关 键 词：卵巢癌 GNAS-AS1 miR-2116-3p 细胞增殖 迁移 侵袭 

分 类 号：R737[医药卫生—肿瘤]