miR-182-5p对宫颈癌细胞增殖和迁移的影响及分子机制  

Effect and molecular mechanisms of miR-182-5p on proliferation and migration of cervical cancer cells

作　　者：胡晓舒 李川 王娜[1] 温一阳[2] HU Xiao-shu;LI Chuan;WANG Na;WEN Yi-yang(The Fifth Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou People's Hospital,Zhengzhou 450003,China;Department of Oncology,Henan Provincial People's Hospital,Zhengzhou 450003,China)

机构地区：[1]河南中医药大学第五临床医学院,郑州人民医院,郑州450003 [2]河南省人民医院肿瘤中心,郑州450003

出　　处：《医药论坛杂志》2024年第16期1681-1685,1691,共6页Journal of Medical Forum

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20230717)。

摘　　要：目的探讨miR-182-5p对宫颈癌细胞的生物学特性影响及其机制。方法选取2019年1月至2022年1月郑州人民医院收集的宫颈癌和癌旁组织样本,运用荧光定量PCR分析miR-182-5p在上述组织中的表达。用携带空载体和miR-182-5p的慢病毒分别感染宫颈癌Hela细胞,构建微小RNA(mircoRNA,miRNA)对照(miR-control组)和miR-182-5p过表达细胞系(miR-182-5p组)。采用5-乙炔基-2'脱氧尿嘧啶核苷(5-ethynyl-2'-deoxyuridine,EdU)染色和细胞计数试剂盒(cell counting kit-8,CCK8)分析两组细胞的增殖能力,划痕实验和Transwell分析两组细胞的迁移能力。采用生信分析和双荧光素酶报告基因检测miR-182-5p的靶基因,并用Western blot验证靶基因的表达。结果宫颈癌组织中miR-182-5p表达水平(0.31±0.02)明显低于癌旁组织(1.01±0.11),差异有统计学意义(t=11.105,P<0.05)。miR-182-5p组细胞miR-182-5p表达水平(2.77±0.43)明显高于miR-control组(1.11±0.09),差异有统计学意义(t=11.963,P<0.05)。miR-182-5p组的细胞增殖能力(1.23±0.05)明显低于miR-control组(1.93±0.07),差异有统计学意义(t=14.771,P<0.05)。miR-182-5p组细胞EdU染色阳性率(20.33±1.63)%明显低于miR-control组(40.32±1.86)%,差异有统计学意义(t=13.997,P<0.05)。miR-182-5p组迁移细胞数量(56.18±4.22)明显低于miR-control组(157.63±8.25),差异有统计学意义(t=18.958,P<0.05)。miR-182-5p组细胞划痕愈合度(21.39±3.09)%明显低于miR-control组(76.17±4.22)%,差异有统计学意义(t=18.145,P<0.05)。生信和双荧光素酶基因证实Rab27A为miR-182-5p的靶基因。miR-182-5p组细胞中Rab27A蛋白表达水平(2.18±0.19)明显高于miR-control组(1.14±0.11),差异有统计学意义(t=8.327,P<0.05)。结论miR-183-5p通过上调Rab27抑制宫颈癌细胞的增殖和迁移。Objective To investigate the effect of miR-182-5p on the biological characteristics of cervical cancer cells and its mechanism.Methods Cervical cancer and paracancer tissue samples collected in Zhengzhou People's Hospital from January 2019 to January 2022 were selected,and the expression of miR-182-5p in the above tissues was analyzed by fluorescence quantitative PCR.Lentiviruses carrying empty vectors and miR-182-5p were infected with cervical cancer Hela cells,respectively,and miRNA control(miR-control group)and miR-182-5p overexpression cell lines(miR-182-5p group)were constructed.5-ethynyl-2'-deoxyuridine(EdU)staining and cell counting kit-8(CCK8)were used to analyze the proliferation ability of two group cells,and scratch and Transwell were used to analyze the migration ability of two group cells.The target gene of miR-182-5p was detected by bioinformatics and dual luciferase reporter gene,and the expression of the target gene was verified by Western blot.Results The expression level of miR-182-5p in cervical cancer tissues(0.31±0.02)was significantly lower than that in paracancer tissues(1.01±0.11,t=11.105,P<0.05).The expression level of miR-182-5p in miR-182-5p group(2.77±0.43)was significantly higher than that in miR-control group(1.11±0.09,t=11.963,P<0.05).The cell proliferation ability of miR-182-5p group(1.23±0.05)was significantly lower than that of miR-control group(1.93±0.07,t=14.771,P<0.05).The positive rate of EdU staining in miR-182-5p group(20.33±1.63)%was significantly lower than that in miR-control group(40.32±1.86)%,and the difference was statistically significant(t=13.997,P<0.05).The number of migrated cells in miR-182-5p group(56.18±4.22)was significantly lower than that in miR-control group[(157.63±8.25),t=18.958,P<0.05].The cell scratch healing rate of miR-182-5p group(21.39±3.09)%was significantly lower than that of miR-control group[(76.17±4.22)%,t=18.145,P<0.05].Rab27A was identified as the target gene of miR-182-5p by bioinformatics and dual luciferase genes.The Rab27A pro

关 键 词：miR-182-5p 宫颈癌 增殖 迁移 

分 类 号：R737.33[医药卫生—肿瘤]