lncRNA GABPB1-IT1靶向下调miR-501抑制宫颈癌细胞增殖、侵袭和迁移的机制  

lncRNA GABPB1-IT1 inhibits the proliferation,invasion and migration of cervical cancer cells by targeting miR-501

作　　者：赵丹华[1] 田艳杰[1] 李秋爽[1] 刘丽[1] 李翠英[1] 孙静莉 Zhao Danhua;Tian Yanjie;Li Qiushuang;Liu Li;Li Cuiying;Sun Jingli(Department of Obstetrics and Gynecology,Tieling Maternal and Infant Hospital,Liaoning 112000,China;Department of Obstetrics and Gynecology,General Hospital of NorthernWar Zone,Liaoning 110016,China)

机构地区：[1]铁岭市妇婴医院妇产科,铁岭112000 [2]北部战区总医院妇产科,沈阳110016

出　　处：《广西医科大学学报》2021年第7期1276-1282,共7页Journal of Guangxi Medical University

基　　金：辽宁省自然科学基金计划重点项目(No.20170540909)。

摘　　要：目的:探讨lncRNA GABPB1-IT1靶向下调miR-501抑制宫颈癌细胞增殖、侵袭和迁移的机制。方法:qRT-PCR方法检测宫颈癌HeLa、Caski、SiHa细胞和正常宫颈上皮Ect1/E6E7细胞中GABPB1-IT1表达水平。宫颈癌SiHa细胞分成Control组、Vector组(转染pcDNA)、GABPB1-IT1组(转染pcDNA-GABPB1-IT1)、GABPB1-IT1+miR-NC组(转染pcDNA-GABPB1-IT1和mimics control)、GABPB1-IT1+miR-501组(转染pcDNA-GABPB1-IT1和miR-501 mimics)。CCK-8法检测细胞增殖;Transwell小室检测细胞迁移和侵袭;Western blotting检测细胞中MMP-2、MMP-9、N-cadherin、E-cadherin蛋白表达。利用starbase数据库对GABPB1-IT1的靶基因进行分析,通过荧光素酶报告系统鉴定靶向关系。结果:宫颈癌HeLa、Caski、SiHa细胞中GABPB1-IT1表达水平低于正常宫颈上皮Ect1/E6E7细胞(均P<0.05)。宫颈癌HeLa、Caski细胞中GABPB1-IT1表达水平高于宫颈癌SiHa细胞(均P<0.05)。与Control组、Vector组比较,GABPB1-IT1组宫颈癌SiHa细胞存活率降低,细胞迁移和侵袭数目减少,细胞中MMP-2、MMP-9、N-cadherin蛋白表达水平降低,E-cadherin蛋白表达水平升高(均P<0.05)。与GABPB1-IT1+miR-NC组比较,GABPB1-IT1+miR-501组宫颈癌SiHa细胞中miR-501水平升高,细胞存活率、细胞迁移数目和侵袭数目均升高,细胞中MMP-2、MMP-9、N-cadherin蛋白表达水平升高,E-cadherin蛋白表达水平降低(均P<0.05)。GABPB1-IT1靶向下调miR-501表达水平。结论:lncRNA GABPB1-IT1靶向下调miR-501抑制宫颈癌细胞增殖、侵袭和迁移。Objective To investigate the mechanism of lncRNA GABPB1-IT1 on the inhibition of proliferation,invasion and migration of cervical cancer cells bytargeting miR-501.Methods:The qRT-PCR method was used to detect the expression level of GABPB1-IT1 in cervical cancer HeLa,Caski,SiHa cells and normal cervical epithelial Ect1/E6E7 cells.Cervical cancer SiHa cells were divided into Control,Vector(transfected with pcDNA),GABPB1-IT1(transfected with pcDNA-GABPB1-IT1),GABPB1-IT1+miR-NC(transfected with pcDNA-GABPB1-IT1 and mimics control),GABPB1-IT1+miR-501 group(transfected with pcDNA-GABPB1-IT1 and miR-501 mimics).CCK-8 method was used to detect cell proliferation.Transwell chamber method was used to detect cell migration and invasion.Western blot method was used to detect MMP-2,MMP-9,N-cadherin,and E-cadherin protein expression.The target genes of GABPB1-IT1 were analyzed using Starbase database,and conformed by luciferase reporter system.Results:The expression level of GABPB1-IT1 in cervical cancer HeLa,Caski and SiHa cells were lower than that in normal cervical epithelial Ect1/E6E7 cells(P<0.05).The expression level of GABPB1-IT1 in cervical cancer HeLa and Caski cells were higher than that in cervical cancer SiHa cells(P<0.05).Compared with the Control and Vector groups,the survival rate of cervical cancer SiHa cells,the migration and invasion cell number,the expression levels of MMP-2,MMP-9,and N-cadherin proteins were decreased,while the expression level of E-cadherin proteins was increased in the GABPB1-IT1 group(P<0.05).Compared with the GABPB1-IT1+miR-NC group,the miR-501 expression,cell survival rate,cell migration and invasion rate,and the expressions of MMP-2,MMP-9,and N-cadherin protein were increased,whereas the E-cadherin protein expression was decreased in GABPB1-IT1+miR-501 group(P<0.05).GABPB1-IT1 targeted and down-regulated miR-501 expression.Conclusion:lncRNA GABPB1-IT1 targeted and downregulated miR-501 to inhibit the proliferation,invasion and migration of cervical cancer cells.

关 键 词：宫颈癌 GA结合蛋白转录因子亚基β1-内含子转录本1 微小RNA(miR)-501 侵袭 迁移 

分 类 号：R737.33[医药卫生—肿瘤]