circ_0010423通过吸附miR-423-5p和miR-3184-5p上调COL1A1对宫颈癌恶性生物学行为的影响  

The effect of circ_0010423 on malignant biological behavior of cervical cancer cells by upregulation of COL1A1 through sponging miR-423-5p and miR-3184-5p

作　　者：毛雪宝 王秀虹 邱彬 李美灵 马丹 蒋金朋 MAO Xuebao;WANG Xiuhong;QIU Bin;LI Meiling;MA Dan;JIANG Jinpeng(Department of Gynecology,the Center Hospital of Xianning City,the First Affiliated Hospital of Hubei College of Science and Technology,Hubei Xianning 437100,China)

机构地区：[1]咸宁市中心医院(湖北科技学院附属第一医院)妇科,湖北咸宁437100

出　　处：《现代肿瘤医学》2024年第11期2001-2009,共9页Journal of Modern Oncology

基　　金：湖北省咸宁市自然科学基金资助项目(编号:2021ZRKX045)。

摘　　要：目的:探讨环状RNA circ_0010423对宫颈癌细胞增殖、侵袭、迁移和上皮间充质转化(epithelial-mesenchymal transition,EMT)的影响及机制。方法:采用qRT-PCR检测40对宫颈癌组织与癌旁组织、5种宫颈癌细胞株(HeLa、ME180、CaSki、HCC94和SiHa)与正常宫颈上皮细胞株(End1/E6E7)中circ_0010423、miR-423-5p、miR-3184-5p及Ⅰ型胶原α1(collagen type I alpha 1 chain,COL1A1)的表达;SiHa细胞株中转染sh-NC(sh-NC组)、sh-circ_0010423(sh-circ_0010423组)、sh-circ_0010423+control vector(sh-circ_0010423+control vector组)及sh-circ_0010423+COL1A1 vector(sh-circ_0010423+COL1A1 vector组),CCK-8实验检测细胞增殖,Transwell侵袭和迁移实验检测细胞侵袭和迁移,Western blot法检测EMT标记物(E-cadherin、N-cadherin及Vimentin蛋白)表达;通过生信分析、荧光素酶及RIP实验验证circ_0010423与miR-423-5p、miR-3184-5p的靶向关系,miR-423-5p、miR-3184-5p与COL1A1的靶向关系。结果:与癌旁组织和正常宫颈上皮细胞株比较,宫颈癌组织和细胞株中circ_0010423、COL1A1表达水平升高,miR-423-5p、miR-3184-5p表达水平降低(P<0.05)。与sh-NC组比较,sh-circ_0010423组SiHa细胞24 h、48 h、72 h OD值、侵袭、迁移细胞数、N-cadherin、Vimentin蛋白表达水平均降低,E-cadherin蛋白表达升高(P<0.05)。COL1A1是miR-423-5p、miR-3184-5p靶基因,circ_0010423作为miR-423-5p、miR-3184-5p分子海绵解除对COL1A1的抑制作用。与sh-circ_0010423+control vector组比较,sh-circ_0010423+COL1A1 vector组SiHa细胞24 h、48 h、72 h OD值、侵袭、迁移细胞数、N-cadherin、Vimentin蛋白表达水平均升高,E-cadherin蛋白表达降低(P<0.05)。结论:circ_0010423在宫颈癌组织和细胞中表达上调,并可以通过吸附miR-423-5p和miR-3184-5p上调COL1A1表达,促进SiHa细胞增殖、侵袭、迁移及EMT,导致宫颈癌发展。Objective:To investigate the effect and mechanism of circRNA circ_0010423 on the proliferation,invasion,migration and epithelial-mesenchymal transition(EMT)of cervical cancer cells.Methods:qRT-PCR was used to detect the expression of circ_0010423,miR-423-5p,miR-3184-5p and collagen typeⅠalpha 1 chain(COL1A1)in 40 pairs of cervical cancer tissues and corresponding adjacent tissues,five cervical cancer cell lines(HeLa,ME180,CaSki,HCC94 and SiHa)and a normal cervical epithelial cell line(End1/E6E7).SiHa cell line was transfected with sh-NC(sh-NC group),sh-circ_0010423(sh-circ_0010423 group),sh-circ_0010423+control vector(sh-circ_0010423+control vector group)and sh-circ_0010423+COL1A1 vector(sh-circ_0010423+COL1A1 vector group),cell proliferation was detected by CCK-8 experiment,cell invasion and migration were detected by Transwell invasion and migration experiment,and the expression of EMT marker(E-cadherin,N-cadherin and Vimentin protein)was detected by Western blot.Bioinformatics analysis,dual luciferase assay and RIP experiment were used to analyze the targeting relationship between circ_0010423,miR-423-5p and miR-3184-5p,and the targeting relationship between miR-423-5p,miR-3184-5p and COL1A1.Results:Compared with adjacent tissues and normal cervical epithelial cell line,cervical cancer tissues and cell lines showed increased expression levels of circ_0010423 and COL1A1,and reduced expression levels of miR-423-5p and miR-3184-5p(P<0.05).Compared with the sh-NC group,the OD value of SiHa cells in the sh-circ_0010423 group at 24 h,48 h and 72 h,the number of invasive and migratory cells,and the expression levels of N-cadherin and Vimentin protein were decreased,while E-cadherin protein expression was increased(P<0.05).COL1A1 is a target gene of miR-423-5p and miR-3184-5p.circ_0010423 act as a molecular sponge of miR-423-5p and miR-3184-5p to deregulate the inhibitory effect of COL1A1.Compared with the sh-circ_0010423+control vector group,the OD value of SiHa cells in the sh-circ_0010423+COL1A1 vector group at 2

关 键 词：circ_0010423 宫颈癌细胞 miR-423-5p miR-3184-5p COL1A1 

分 类 号：R737.33[医药卫生—肿瘤]