LncRNA SNHG1通过miR-101-3p/MYLIP轴调节宫颈癌细胞增殖、凋亡和迁移的研究  

LncRNA SNHG1 regulates the proliferation,apoptosis and migration of cervical cancer cells through the miR-101-3p/MYLIP axis

作　　者：白小英[1] 曹蕾[1] 张卫霞[1] 陈琳[1] 胡海燕[1] 杨婷[1] Bai Xiaoying;Cao Lei;Zhang Weixia;Chen Lin;Hu Haiyan;Yang Ting(Department of Gynecology,the Second Affiliated Hospital of Xi'an Medical College,Xi an Shaanxi 710038,P.R.China)

机构地区：[1]西安医学院第二附属医院妇科,陕西西安710038

出　　处：《中国计划生育和妇产科》2023年第4期38-43,60,I0001,共8页Chinese Journal of Family Planning & Gynecotokology

基　　金：陕西省重点研发计划(项目编号:2019SF-051)。

摘　　要：目的 探究长链非编码RNA(lncRNA)小核仁RNA宿主基因1(SNHG1)对宫颈癌(cervical cancer, CC)细胞增殖、迁移和侵袭的影响及分子机制。方法 实时荧光定量PCR(qRT-PCR)和Western blot检测人正常宫颈上皮细胞(H8)和人CC细胞(Hela、Caski、C-33A、SiHa)中SNHG1、microRNA-101-3p(miR-101-3p)、肌球蛋白调节轻链相互作用蛋白(MYLIP)的mRNA和蛋白表达。体外培养Hela细胞,使用Lipofectamine 3000试剂将si-SNHG1、anti-miR-101-3p及其阴性对照si-NC、inhibitor-NC转染到Hela细胞中,分为对照(Control)组、si-NC组、si-SNHG1组、si-SNHG1+anti-NC组、si-SNHG1+anti-miR-101-3p组。转染48 h后,qRT-PCR检测细胞中SNHG1、miR-101-3p和MYLIP mRNA的表达水平,Western blot检测细胞中MYLIP蛋白表达;CCK-8法检测细胞增殖活力,流式细胞术检测细胞凋亡,Transwell小室实验检测细胞迁移、侵袭能力;通过双荧光素酶报告(DLRA)和RNA结合蛋白免疫沉淀(RIP)实验验证Hela细胞中SNHG1、miR-101-3p和MYLIP的调控作用。结果 CC细胞系(Hela、Caski、C-33A、SiHa)中SNHG1、MYLIP mRNA和蛋白水平升高,miR-101-3p表达水平降低(P<0.05)。敲低SNHG1可显著上调miR-101-3p表达,降低MYLIP表达,抑制Hela细胞增殖、迁移和侵袭,并促进Hela细胞凋亡(均P<0.05);下调miR-101-3p可显著减弱SNHG1敲低的促凋亡和抗迁移、侵袭作用(均P<0.05)。DLRA和RIP实验证实SNHG1可以靶向并结合Hela细胞中的miR-101-3p, MYLIP是miR-101-3p的靶标。结论 SNHG1敲低可能是通过上调miR-101-3p来抑制MYLIP表达,进而抑制CC细胞增殖、迁移和侵袭,并促进CC细胞凋亡。Objective To investigate the impacts and molecular mechanism of long non-coding RNA(lncRNA)small nucleolar RNA host gene 1(SNHG1)on the proliferation,migration and invasion of cervical cancer(CC)cells.Methods Quantitative realtime PCR(qRT-PCR)and Western blot were performed to detect the mRNA and protein expression of SNHG1,microRNA-101-3p(miR-101-3p),myosin regulated light chain interacting protein(MYLIP)in human normal cervical epithelial cells(H8)and human CC cells(Hela,Caski,C-33A,SiHa).Hela cells were cultured in vitro,and the si-SNHG1,anti-miR-101-3p and their negative controls si-NC and inhibitor-NC were transfected into Hela cells by Lipofectamine 3000 reagent,and they were grouped into Control group,si-NC group,si-SNHGI group,si-SNHG1+anti-NC group,and si-SNHGI+anti-miR-101-3p group.After 48 hours of transfection,the expression levels of SNHG1,miR-101-3p and MYLIP mRNA in cells were detected by quantitative real-time PCR(qRT-PCR),Western blot was performed to detect the protein expression of MYLIP in cells;the cell proliferation activity was detected by CCK-8 method,the apoptosis was detected by flow cytometry,Transwell chamber assay was performed to detect cell migration and invasion abilities;the regulatory roles of SNHG1,miR-101-3p and MYLIP in Hela cells were verified by dual luciferase reporter assay(DLRA)and RNA binding protein immunoprecipitation(RIP)experiments.Results The mRNA and protein levels of SNHG1 and MYLIP in CC cell lines(Hela,Caski,C-33A,SiHa)were increased,and the expression level of miR-101-3p was decreased(P<0.05).Knockdown of SNHG1 significantly up-regulated the expression of miR-101-3p,decreased the expression of MYLIP,inhibited the proliferation,migration and invasion of Hela cells,and promoted the apoptosis of Hela cells(all P<0.05);down-regulation of miR-101-3p could significantly attenuate the pro-apoptotic,anti-migration and invasion effects of SNHG1 knockdown(all P<0.05).DLRA and RIP experiments confirmed that SNHG1 could target and bind to miR-101-3p in Hela cells,and MYLIP

关 键 词：宫颈癌 长链非编码RNA小核仁RNA宿主基因1 microRNA-101-3p 肌球蛋白调节轻链相互作用蛋白 增殖 凋亡 迁移 

分 类 号：R711.74[医药卫生—妇产科学]