微小RNA-149调控肿瘤细胞增殖和转移的分子机制  

Molecular mechanism of microRNA-149 regulating proliferation and metastasis of ovarian cancer cells

作　　者：孙桂霞[1] 张冬丽[1] 杨少琴[1] 曹芹雪[1] 田君[1] Sun Guixia;Zhang Dongli;Yang Shaoqin;Cao Qinxue;Tian Jun(Department of Gynaecology,the Huaihe Hospital of Henan University,Kaifeng 475000,China)

机构地区：[1]河南大学淮河医院妇瘤科,开封475000

出　　处：《中华实验外科杂志》2023年第9期1816-1819,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划项目(LHGJ20210574)。

摘　　要：目的探讨微小RNA(miRNA,miR)-149调控卵巢癌细胞增殖和转移的影响及其分子机制。方法选取2019年1月至2021年1月河南大学淮河医院手术切除的47例卵巢癌组织和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析肿瘤组织和癌旁组织中miR-149水平。并采用荧光定量PCR分析人卵巢上皮细胞HOSEpiC、卵巢癌细胞SW620和skov3中miR-149水平;采用miRNA和miR-149慢病毒感染SW620细胞,命名为miRNA组和miR-149组,采用细胞计数试剂盒(CCK-8)分析两组细胞的增殖能力;采用流式细胞术分析两组细胞凋亡和周期变化;采用Transwell分析两组细胞的迁移能力;生物信息学和双荧光素酶报告基因分析miR-149的靶基因;采用蛋白质印迹法(Western blot)分析miR-149靶蛋白表达、增殖、凋亡、周期和迁移相关蛋白表达水平。组间计量数据比较采用t检验。结果癌旁组织miR-149表达水平(1.07±0.15)明显高于卵巢癌组织(0.73±0.09),差异有统计学意义(t=13.490,P<0.05)。人卵巢上皮细胞HOSEpiC miR-149表达水平(1.27±0.10)明显高于卵巢癌细胞SW620和skov3(0.80±0.08、0.70±0.08),差异有统计学意义(t=8.859、10.740,P<0.05)。miRNA对照组细胞48 h吸光度值(1.91±0.08)明显高于miR-149组(1.23±0.18),差异有统计学意义(t=8.655,P<0.05)。miRNA对照组细胞凋亡比例[(3.69±1.06)%]明显低于miR-149组[(18.90±2.35)%],差异有统计学意义(t=14.470,P<0.05)。miRNA对照组细胞G0/G1期比例[(39.91±2.86)%]明显高于miR-149组[(31.33±2.42)%],差异有统计学意义(t=5.616,P<0.05)。miRNA对照组细胞S期比例[(16.73±1.51)%]明显低于miR-149组[(20.48±2.04)%],差异有统计学意义(t=3.619,P<0.05)。miRNA对照组细胞凋亡比例[(127.67±21.69)个]明显高于miR-149组[(99.50±10.21)个],差异有统计学意义(t=2.887,P<0.05)。miRNA对照组细胞细胞核增殖抗原(Ki-67)和黏着斑激酶(FAK)蛋白表达水平(0.94±0.06、0.91±0.05)明显高于miR-149组(0.73±0.06、0.59±Objective To investigate the effect of microRNA(miR)-149 on the proliferation and metastasis of ovarian cancer cells and its molecular mechanism.Methods Totally,47 cases of ovarian cancer tissues and adjacent tissues resected in our hospital from January 2019 to January 2021 were selected as research objects.The miR-149 level in tumor tissues and adjacent tissues was analyzed by fluorescent quantitative polymerase chain reaction(PCR).The levels of miR-149 in human ovarian epithelial cells HOSEpiC,ovarian cancer cells SW620 and skov3 were analyzed by fluorescent quantitative PCR.SW620 cells were infected with miRNA and miR-149 lentivirus,which were named miRNA group and miR-149 group respectively.The proliferation ability of the two groups cells was analyzed by cell counting kit-8(CCK-8).Apoptosis and cell cycle were analyzed by flow cytometry.The migration ability of the two groups of cells was analyzed by Transwell.Bioinformatics and double luciferase reporter gene were applied to analyze the target gene of miR-149.The expression levels of miR-149 target protein,proliferation,apoptosis,cycle and migration related proteins were analyzed by Western blotting.The measurement data between groups were compared by t test.Results The expression level of miR-149 in adjacent tissues(1.07±0.15)was significantly higher than that in ovarian cancer tissues(0.73±0.09,t=13.490,P<0.05).The expression level of HOSEpiC miR-149 in human ovarian epithelial cells(1.27±0.10)was significantly higher than that in ovarian cancer cells SW620 and skov3(0.80±0.08,0.70±0.08,t=8.859,10.740,P<0.05).The 48 h absorbance(A)value in the miRNA control group(1.91±0.08)was significantly higher than that in the miR-149 group(1.23±0.18,t=8.655,P<0.05).The percentage of apoptosis in the miRNA control group[(3.69±1.06)%]was significantly lower than that in the miR-149 group[(18.90±2.35)%,t=14.470,P<0.05].The proportion of G 0/G1 phase cells in the miRNA control group[(39.91±2.86)%]was significantly higher than that in the miR-149 group[(31.33±

关 键 词：微小RNA 卵巢癌 增殖 细胞周期 凋亡 转移 

分 类 号：R730.2[医药卫生—肿瘤]