miR-3074通过靶向EPS8基因对宫颈癌细胞增殖和侵袭的影响  

Effects of miR-3074 on Proliferation and Invasion of Cervical Cancer Cells by Targeting EPS8gene

作　　者：罗全慧 尹恒 黄柯 王锦星 赵琳[1] LUO Quanhui;YIN Heng;HUANG Ke(Obstetrics and Gynecology Center,Taihe Hospital Affiliated to Hubei Medical College,Hubei 442000,China)

机构地区：[1]湖北医药学院附属太和医院妇产中心,十堰442000

出　　处：《医学研究杂志》2023年第1期66-71,共6页Journal of Medical Research

基　　金：湖北省科技计划项目(2021CFB162)。

摘　　要：目的探讨miR-3074通过靶向调节表皮生长因子受体通路底物8(epidermal growth factor receptor substrate 8,EPS8)基因表达对宫颈癌细胞增殖和侵袭的影响。方法应用OncoLnc在线数据库分析miR-3074表达水平与宫颈癌患者总生存期的关系。实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)检测miR-3074在宫颈癌细胞系(SiHa、HCC94、C33A、HeLa)和正常宫颈上皮细胞H8中的表达。选取miR-3074表达最低的细胞系,分别转染对照模拟物(对照组)和miR-3074模拟物(miR-3074组)。CCK-8法和Transwell实验分别检测细胞增殖活力和侵袭能力。生物信息学软件RNAhybrid预测miR-3074的靶基因,双荧光素酶报告基因实验验证miR-3074与靶基因的靶向关系。qRT-PCR和Western blot法检测miR-3074对靶基因表达的影响。结果miR-3074表达水平较高的宫颈癌患者总生存期长于miR-3074表达水平较低的患者,差异有统计学意义(P<0.01)。与H8细胞比较,宫颈癌细胞系中miR-3074表达降低(P<0.05),miR-3074表达最低的细胞系是HCC94(P<0.01)。对照组和miR-3074组miR-3074表达分别为1.03±0.17和13.49±2.66,差异有统计学意义(P<0.01)。与对照组比较,miR-3074组HCC94细胞增殖活力降低(P<0.05),细胞侵袭能力降低(P<0.01)。miR-3074能作用于EPS8mRNA的3′非翻译区(P<0.01)。与对照组比较,miR-3074组EPS8基因表达显著降低(P<0.01)。结论miR-3074在宫颈癌细胞系中表达下调,miR-3074能够抑制宫颈癌HCC94细胞的增殖和侵袭,其机制可能与靶向抑制EPS8表达有关。Objective To explore the effect of miR-3074 on the proliferation and invasion of cervical cancer cell through targeted regulation of epidermal growth factor receptor substrate 8(EPS8)gene expression.Methods The OncoLnc online database was used to analyze the relationship between the expression level of miR-3074 and the overall survival of patients with cervical cancer.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-3074 in cervical cancer cell lines(SiHa,HCC94,C33A,HeLa)and normal cervical epithelial cells H8.The cell line with the lowest expression of miR-3074 was selected and transfected with the control mimics(control group)and miR-3074mimics(miR-3074 group).CCK-8 assay and Transwell assay were used to detect cell proliferation activity and invasion ability.The bioinformatics software RNAhybrid was used to predict the target gene of miR-3074,and the dual-luciferase reporter gene experiment was used to verify the targeting relationship between miR-3074 and the target gene.RT-qPCR and Western blot were used to detect the effect of miR-3074 on target gene expression.Results The overall survival of cervical cancer patients with higher miR-3074 expression level was longer than that of patients with lower miR-3074 expression level,and the difference was statistically significant(P<0.01).Compared with H8 cells,the expression of miR-3074 in cervical cancer cell lines was reduced(P<0.05),and the cell line with the lowest expression of miR-3074 was HCC94(P<0.01).The expression of miR-3074 in the control group and miR-3074 group was 1.03±0.17 and 13.49±2.66,respectively,and the difference was statistically significant(P<0.01).Compared with the control group,the proliferation activity of HCC94 cells in miR-3074 group was decreased(P<0.05),and cell invasion ability was decreased(P<0.01).miR-3074 can bind to the 3′untranslated region of EPS8mRNA(P<0.01).Compared with the control group,the expression of EPS8gene in miR-3074group was significantly decreased(P<

关 键 词：宫颈癌 微小RNA 表皮生长因子受体通路底物8 细胞增殖 细胞侵袭 

分 类 号：R737.33[医药卫生—肿瘤]