LncRNA RP11-499E18.1对卵巢癌细胞恶性生物学行为的影响  

Effects of lncRNA RP11-499E18.1 on the malignant biological behavior of ovarian cancer cells

作　　者：李瑛花 杨娟 LI Yinghua;YANG Juan(Gynecologic Oncology Ward III,Hunan Cancer Hospital,Affiliated Cancer Hospital of Xiangya School of Medicine,Central South University,Changsha 410013;Gynecologic Oncology Ward V,Hunan Cancer Hospital,Affiliated Cancer Hospital of Xiangya School of Medicine,Central South University,Changsha 410013,China)

机构地区：[1]中南大学湘雅医学院附属肿瘤医院/湖南省肿瘤医院妇瘤二科,长沙410013 [2]中南大学湘雅医学院附属肿瘤医院/湖南省肿瘤医院妇瘤五科,长沙410013

出　　处：《中南大学学报(医学版)》2025年第1期1-10,共10页Journal of Central South University :Medical Science

基　　金：湖南省自然科学基金(2021JJ40322);湖南省科技创新计划项目(2020SK51102)。

摘　　要：目的:卵巢癌是常见的妇科恶性肿瘤,晚期患者的预后较差。本研究筛选卵巢癌预后相关的差异表达长链非编码RNA(long noncoding RNAs,lncRNA),并探究其对卵巢癌细胞恶性生物学行为的影响。方法:从基因表达综合(Gene Expression Omnibus,GEO)数据库中获取卵巢癌相关lncRNAs数据集,并采用lncRNA测序结合GEO数据库中相关临床信息筛选潜在的肿瘤抑制lncRNA。采用反转录聚合酶链反应(reverse transcription PCR,RT-PCR)检测lncRNA RP11-499E18.1在卵巢组织及相应的癌旁组织、卵巢正常上皮细胞系IOSE80及卵巢癌细胞系中的表达情况;采用荧光原位杂交(fluorescence in situ hybridization,FISH)技术检测lncRNA RP11-499E18.1在卵巢癌细胞中的定位。将卵巢癌细胞系CaOV3和SKOV3分为3组:阴性对照(negative control,NC)组、敲减(si-RP11-499E18.1)组和过表达(pcDNA-RP11-499E18.1)组。采用四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)实验和Transwell实验分别检测lncRNA RP11-499E18.1对卵巢癌细胞增殖和迁移的影响;采用蛋白质印迹法检测lncRNA RP11-499E18.1对卵巢癌细胞上皮-间充质转化(epithelial-mesenchymal transition,EMT)相关分子表达的影响。将BALB/c裸鼠小鼠分为实验组(注射转染pcDNA-RP11-499E18.1的CaOV3细胞)和对照组(注射转染空载体pcDNA的CaOV3细胞)。采用免疫组织化学法检测实验组和对照组卵巢癌组织中Caspase 3和Ki67的表达情况。结果:LncRNA测序结果显示lncRNA RP11-499E18.1为与卵巢癌预后相关的差异表达lncRNA。对GEO数据库中相关临床数据进行分析,结果显示:与lncRNA RP11-499E18.1高表达的卵巢癌患者相比,lncRNA RP11-499E18.1低表达的卵巢癌患者的总生存期、无进展生存期更短(均P<0.05)。LncRNA RP11-499E18.1为潜在的肿瘤抑制lncRNA。LncRNA RP11-499E18.1在卵巢癌组织的表达明显低于癌旁组织(P<0.001),在卵巢癌细胞系CaOV3、OVCAR3、SKOV3、A2780中的表达显著低于IOSE80(均P<0.001),�Objective:Ovarian cancer is a common gynecologic malignancy,with poor prognosis in advanced stages.This study aimed to identify differentially expressed long noncoding RNA(lncRNA)associated with ovarian cancer prognosis and to explore the effects of lncRNA RP11-499E18.1 on the malignant biological behavior of ovarian cancer cells.Methods:Ovarian cancer-related lncRNA datasets were obtained from the Gene Expression Omnibus(GEO)database.Differentially expressed and prognostically relevant tumor-suppressive lncRNAs were screened using lncRNA sequencing combined with clinical data.Reverse transcription PCR(RT-PCR)was used to detect the expression of lncRNA RP11-499E18.1 in ovarian cancer tissues,adjacent normal tissues,the IOSE80 normal ovarian epithelial cell line,and various ovarian cancer cell lines.Fluorescence in situ hybridization(FISH)was performed to determine its subcellular localization.Ovarian cancer cell lines CaOV3 and SKOV3 were divided into 3 groups:a negative control(NC)group,a knockdown(si-RP11-499E18.1)group,and a overexpression(pcDNA-RP11-499E18.1)group.Methyl thiazolyl tetrazolium(MTT)and Transwell assays were used to assess the effects of lncRNA RP11-499E18.1 on cell proliferation and migration.Western blotting was used to evaluate its effect on epithelial-mesenchymal transition(EMT)-related molecules.BALB/c nude mice were injected with CaOV3 cells transfected with pcDNARP11-499E18.1(experimental group)or empty vector(control group),and tumor growth was monitored.Immunohistochemistry was used to assess the expression of Caspase 3 and Ki67 in tumor tissues.Results:LncRNA sequencing identified RP11-499E18.1 as a differentially expressed and associated with prognosis.GEO data analysis showed that low RP11-499E18.1 expression was correlated with shorter overall and progression-free survival(both P<0.05).Its expression was significantly lower in ovarian cancer tissues and cell lines compared to normal controls(P<0.05 or P<0.001),and it was localized in both the nucleus and cytoplasm.In CaOV3 and SKOV3

关 键 词：卵巢癌 长链非编码RNA RP11-499E18.1 上皮-间充质转化 恶性生物学行为 

分 类 号：R737.31[医药卫生—肿瘤]