长链非编码RNA C10orf25靶向miRNA-671-5p对前列腺癌细胞增殖和侵袭能力的影响  

Effects of long non-coding RNA C10orf25 targeting miRNA-671-5p on the proliferation and invasion of prostate cancer cells

作　　者：赵云飞 王小英[1] 谢芳 黄耿[1] 王洪[2] 刘佳[3] Zhao Yunfei;Wang Xiaoying;Xie Fang;Huang Geng;Wang Hong;Liu Jia(Department of Urology,Huangshi Central Hospital,Affiliated Hospital of Hubei Polytechnic University,Huangshi 435000,China;Department of Oncology,Renji Hospital Affiliated to Shanghai Jiao Tong University,Shanghai 518052,China;Department of Ultrasound Imaging,Huangshi Central Hospital,Affiliated Hospital of Hubei Polytechnic University,Huangshi 435000,China)

机构地区：[1]黄石市中心医院(湖北理工学院附属医院)泌尿外科,黄石435000 [2]上海交通大学附属仁济医院肿瘤科,上海518052 [3]黄石市中心医院(湖北理工学院附属医院)超声影像科,黄石435000

出　　处：《肿瘤研究与临床》2024年第7期509-514,共6页Cancer Research and Clinic

基　　金：国家自然科学基金(81802554)。

摘　　要：目的探讨长链非编码RNA(lncRNA)C10orf25对前列腺癌细胞增殖和侵袭能力的影响及miRNA-671-5p(miR-671-5p)在其中可能的作用。方法利用基因表达综合(GEO)数据库中数据(数据更新时间2023年1月),分析C10orf25在137例前列腺癌组织和癌旁组织中表达水平的差异。选择前列腺癌C4-2B、DU-145、22Rv1、PC-3、LNCaP细胞株和永生化前列腺上皮RWPE-1细胞株,应用实时荧光定量聚合酶链反应(qRT-PCR)检测各细胞株C10orf25的相对表达量。选取C10orf25相对表达量最低的22Rv1细胞,分为对照组(转染阴性质粒)和C10orf25组(转染载有C10orf25序列质粒);CCK-8法检测两组22Rv1细胞第1、2、3、4、5天增殖能力(以吸光度值表示);Transwell法检测两组22Rv1细胞的侵袭能力。利用Linc2GO软件预测miR-671-5p与C10orf25具有结合位点,双荧光素酶报告基因实验验证C10orf25与miR-671-5p的靶向关系。qRT-PCR检测两组22Rv1细胞C10orf25和miR-671-5p相对表达量。蛋白质印迹法检测两组22Rv1细胞NF-κB信号通路相关蛋白的表达。结果GEO数据库中,C10orf25在前列腺癌组织中的相对表达量低于癌旁组织(P<0.01)。C10orf25在永生化前列腺上皮细胞株RWPE-1和前列腺癌细胞株C4-2B、DU-145、22Rv1、PC-3、LNCaP中的相对表达量分别为1.00±0.05、0.63±0.04、0.42±0.03、0.18±0.04、0.81±0.02、0.50±0.07,差异有统计学意义(F=43.29,P<0.05)。C10orf25组22Rv1细胞的增殖能力从第2天开始均低于对照组,差异均有统计学意义(均P<0.05)。对照组和C10orf25组侵袭细胞数分别为(97±11)个和(36±9)个,差异有统计学意义(t=4.15,P<0.01)。Linc2GO软件预测结果显示,C10orf25与miR-671-5p具有结合位点。双荧光素酶报告基因实验结果显示,miR-671-5p和C10orf25野生型质粒共转染22Rv1细胞的相对荧光活性低于miR-NC和C10orf25野生型质粒共转染细胞,差异有统计学意义(P<0.01),而miR-671-5p或miR-NC与C10orf25突变型质粒共转染时,22Rv1细胞的相�Objective To explore the effect of long non-coding RNA(lncRNA)C10orf25 on the proliferation and invasion ability of prostate cancer cells and the possible role of miRNA-671-5p(miR-671-5p).Methods Data from the Gene expression omnibus(GEO)database(data updated in January 2023)were used to analyze the differences in the expression levels of C10orf25 in 137 cases of prostate cancer tissues and paracancerous tissues.Prostate cancer C4-2B,DU-145,22Rv1,PC-3,LNCaP cell lines and immortalized prostate epithelial RWPE-1 cell lines were selected,and then real-time quantitative fluorescence polymerase chain reaction(qRT-PCR)was used to detect the relative expression levels of C10orf25 in cell lines.The 22Rv1 cells with the lowest relative expression level of C10orf25 were selected and divided into the control group(transfected with negative plasmid)and the C10orf25 group(transfected with C10orf25 plasmid);the CCK-8 method was used to detect the proliferation activity of 22Rv1 cells in both groups at day 1,2,3,4,5(expressed as absorbance value);the Transwell method was used to detect the invasion ability of 22Rv1 cells.Linc2GO software was used to predict miR-671-5p with binding sites for C10orf25.Dual luciferase reporter gene assay was used to verify the targeting relationship between C10orf25 and miR-671-5p.qRT-PCR was used to detect the relative expression levels of C10orf25 and miR-671-5p.Western blot was used to detect the expression of proteins related to the NF-κB signaling pathway of 22Rv1 cells in the both groups.Results In the GEO database,the relative expression level of C10orf25 in prostate cancer tissues was lower than that in paracancerous tissues(P<0.01).The relative expression levels of C10orf25 in immortalized prostate epithelial cell line RWPE-1 and prostate cancer cell lines C4-2B,DU-145,22Rv1,PC-3,and LNCaP were 1.00±0.05,0.63±0.04,0.42±0.03,0.18±0.04,0.81±0.02,0.50±0.07,and the difference was statistically significant(F=43.29,P<0.05).The proliferation ability of 22Rv1 cells in C10orf25 group was l

关 键 词：前列腺肿瘤 RNA 长链非编码 细胞增殖 细胞侵润 C10orf25 

分 类 号：R737.25[医药卫生—肿瘤]