RP11-79H23.3通过抑制miR-410表达调控前列腺癌发生和发展  

RP11-79H23.3 regulates the development and progression of prostate cancer by inhibiting the expression of miR-410

作　　者：柯芹 毛青[1] 陈小刚[1] 江伟 刘伟伟[2] 柳永 Ke Qin;Mao Qing;Chen Xiaogang;Jiang Wei;Liu Weiwei;Liu Yong(Department of Urology,Huangshi Central Hospital,Edong Healthcare Group,Huangshi 435000,China;Department of Laboratory Medicine,Second Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 310009,China)

机构地区：[1]鄂东医疗集团黄石市中心医院泌尿外科,黄石435000 [2]浙江大学医学院附属第二医院检验科,杭州310009

出　　处：《国际外科学杂志》2024年第11期746-751,F0004,共7页International Journal of Surgery

基　　金：国家自然科学基金(81802571)。

摘　　要：目的探讨长链非编码RNA RP11-79H23.3在前列腺癌发生和发展中的作用机制。方法利用lnCAR数据库分析前列腺癌组织和癌旁组织中RP11-79H23.3的相对表达水平。通过实时荧光定量聚合酶链反应(RT-qPCR)检测前列腺癌细胞系C4-2B、LNCaP、DU-145、22Rv1中RP11-79H23.3的表达水平。以22Rv1为研究目标,利用克隆形成实验及划痕实验检测过表达RP11-79H23.3对22Rv1细胞增殖和迁移的作用。利用LncRNome和lncACTdb数据库预测RP11-79H23.3的下游基因和结合序列。利用癌症基因组图谱(TCGA)数据库分析前列腺癌组织中RP11-79H23.3与miR-410表达的相关性。通过双荧光素酶报告基因实验确认RP11-79H23.3和miR-410的结合靶点。采用RT-qPCR法检测RP11-79H23.3对miR-410表达的影响。采用Western blotting法检测RP11-79H23.3对22Rv1细胞中磷酸酶和张力蛋白同源物/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PTEN/AKT/mTOR)信号通路蛋白表达的影响。计量资料以均数±标准差(±s)表示,两组间比较采用配对样本t检验,多组间比较采用单因素方差分析。结果与癌旁组织相比,RP11-79H23.3在前列腺癌组织中呈低表达(P<0.01)。与正常前列腺上皮细胞RWPE-1相比,RP11-79H23.3在前列腺癌细胞系C4-2B、LNCaP、DU-145、22Rv1中均呈低表达(P<0.05)。对照组和RP11-79H23.3组22Rv1细胞中RP11-79H23.3的相对表达水平分别为1.02±0.30和8.94±1.95,与对照组相比,22Rv1细胞成功过表达RP11-79H23.3(t=4.04,P<0.01)。对照组和RP11-79H23.3组22Rv1细胞克隆形成数分别为(166.10±18.35)个和(35.03±6.98)个,与对照组相比,过表达RP11-79H23.3可抑制22Rv1细胞的增殖(t=6.67,P<0.01)。对照组和RP11-79H23.3组22Rv1细胞迁移率分别为(67.40±6.29)%和(26.42±6.24)%,与对照组相比,过表达RP11-79H23.3可抑制22Rv1细胞迁移(t=5.71,P<0.01)。双荧光素酶报告基因实验结果显示,RP11-79H23.3直接与miR-410结合(t=6.20,P<0.01)。对照组和RP11-79H23.3组22Rv1细胞中miR-410ObjectiveTo explore the mechanism of long non-coding RNA RP11-79H23.3 in the development and progression of prostate cancer.MethodsThe lnCAR database was used to analyze the RP11-79H23.3 content in prostate cancer tissues and adjacent tissues.RP11-79H23.3 content in prostate cancer cell lines C4-2B,LNCaP,DU-145,and 22Rv1 was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Taking 22Rv1 as the research target,colony formation experiments and scratch experiments were used to detect the effects of overexpression of RP11-79H23.3 on the proliferation and migration of 22Rv1 cells.The LncRNome and lncACTdb databases were used to predict the downstream gene and binding sequences of RP11-79H23.3.The Cancer Genome Atlas(TCGA)database was used to analyze the correlation between RP11-79H23.3 and miR-410 expression in prostate cancer tissues.The binding of RP11-79H23.3 and miR-410 was confirmed by dual-luciferase reporter gene experiment.The effect of RP11-79H23.3 on the expression of miR-410 was detected by RT-qPCR.Western blotting was used to detect the effect of RP11-79H23.3 on the expression of phosphatase and tensin homolog/protein kinase B/mammalian target of rapamycin(PTEN/AKT/mTOR)signaling pathway proteins in 22Rv1 cells.The measurement data were expressed as mean±standard deviation(±s),paired sample t-test was used for comparison between two groups,and one-way analysis of variance was used for comparison between multiple groups.ResultsCompared with adjacent tissues,RP11-79H23.3 was lowly expressed in prostate cancer tissues(P<0.01).Compared with normal prostate epithelial cells RWPE-1,RP11-79H23.3 was lowly expressed in prostate cancer cell lines C4-2B,LNCaP,DU-145,and 22Rv1(P<0.05).The expression of RP11-79H23.3 in 22Rv1 cells in the control group and RP11-79H23.3 group were 1.02±0.30 and 8.94±1.95,respectively.22Rv1 cells were successfully overexpressed RP11-79H23.3 compared with the control group(t=4.04,P<0.01).The number of 22Rv1 cell clones in the control group and RP11-79H

关 键 词：前列腺肿瘤 细胞增殖 细胞迁移分析 RP11-79H23.3 miR-410 

分 类 号：R737.25[医药卫生—肿瘤]