Linc01278对前列腺癌细胞生物学行为的影响及机制研究  

Study on the effects and mechanism of Linc01278 on the biological behavior of prostate cancer cells

作　　者：孟然[1] 郑秀霞[2] 朱磊[1] 吴丽娜[3] 王雷[1] Meng Ran;Zheng Xiuxia;Zhu Lei;Wu Lina;Wang Lei(Department of Urology,the First People’s Hospital of Shangqiu,Shangqiu 476100,China;Laboratory of Molecular Biology,the First People’s Hospital of Shangqiu,Shangqiu 476100,China;Department of Endocrinology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]河南省商丘市第一人民医院泌尿外科,476100 [2]河南省商丘市第一人民医院分子生物实验室,476100 [3]郑州大学第一附属医院内分泌科,450014

出　　处：《中华内分泌外科杂志》2021年第6期578-582,共5页Chinese Journal of Endocrine Surgery

基　　金：国家自然科学基金青年科学基金(81700719)。

摘　　要：目的探讨长链非编码RNA(LncRNA)Linc01278对前列腺癌细胞增殖、侵袭迁移、肿瘤表型的影响及分子机制。方法收集2018年12月至2019年12月商丘市第一人民医院泌尿外科确诊的46例前列腺癌患者的肿瘤组织及癌旁组织,实时荧光定量PCR(qPCR)检测组织中Linc01278及miR-145的表达含量。培养人前列腺癌细胞PC-3,采用细胞转染技术将PC-3细胞分为:空白组、对照组、过表达组及恢复组。空白组未予以任何处理,对照组转染空白对照载体及miRNA阴性对照,过表达组转染Linc01278过表达载体及miRNA阴性对照,恢复组转染Linc01278过表达载体及miR-145模拟物(mimics)。双荧光素酶报告基因实验分析Linc01278与miR-145的结合作用。Transwell实验检测各组细胞侵袭迁移能力;CCK-8法检测各组细胞增殖能力。Western blot检测各组细胞中Oct4与Sox2的表达含量;qPCR检测各组细胞中Oct4、Sox2及miR-145的表达含量。结果相比癌旁组织,Linc01278在前列腺癌组织中显著高表达(0.012±0.002 vs 0.022±0.002)(P=0.0072),miR-145则显著低表达(0.117±0.011 vs 0.081±0.007)(P=0.0005),两者表达呈负相关(P=0.0012)。Targetscan分析发现Linc01278转录本804-825位置与miR-145存在碱基互补配对。双荧光素酶报告基因结果显示miR-145 mimics可显著下调Linc01278的荧光素酶活性(P<0.01)。相比空白组及对照组,过表达组侵袭及迁移细胞、相对增殖能力及Oct4与Sox2 mRNA及蛋白表达均显著增高,miR-145表达显著降低(P<0.05);而相比过表达组,恢复组侵袭及迁移细胞、相对增殖能力、Oct4与Sox2 mRNA及蛋白表达均显著降低,miR-145表达显著增高(P<0.01)。结论Linc01278可通过特异性吸附miR-145,从而促进前列腺癌细胞增殖、侵袭及迁移能力。Objective To investigate the effect and molecular mechanism of long non-coding RNA(Linc01278)on the proliferation,invasion and migration,and tumor phenotype of prostate cancer cells.Methods Prostate cancer tissues and corresponding normal tissues adjacent to the cancer were obtained in 46 patients with prostate cancer confirmed by the hospital from Dec.2018 to Dec.2019.Real-time fluorescence quantitative PCR(qPCR)was used to detect the expression levels of Linc01278 and miR-145 in prostate cancer tissues and adjacent normal tissues.The prostate cancer PC-3 cells were transfected with plasmids and cells were divided into four groups:blank group,control group,overexpression group and rescue group.The blank group was not given any treatment;the control group was transfected with blank control vector and miRNA Control;the overexpression group was transfected with Linc01278 overexpression vector and miRNA Control;the rescue group was transfected with Linc01278 overexpression vector and miR-145 mimics.Bioinformatics analysis was used to predict the binding site of Linc01278 to miR-145.Dual Luciferase Reporter Gene Assay was used to analyze the adsorption of Linc01278 on miR-145;Transwell experiment was used to detect the invasion and migration ability of the four groups of cells;CCK-8 method was used to detect the cell proliferation ability of the four groups of cells.Western blot was used to detect the expression of OCT4 and SOX2 in the four groups of cells;qPCR was used to detect the expression of OCT4,SOX2 and miR-145 in the four groups of cells.Results Compared with adjacent normal tissues,Linc01278 was significantly higher in prostate cancer tissues(adjacent normal tissues:0.012±0.002 vs prostate cancer tissues:0.022±0.002,P=0.0072),while miR-145 was significantly lower(adjacent normal tissues:0.117±0.011 vs prostate cancer tissues:0.081±0.007,P=0.0005).Correlation of both was negative significantly(P=0.0012).Targetscan analysis revealed that the 804-825 position of the Linc01278 transcript had a complementary

关 键 词：前列腺癌 Linc01278 MIR-145 OCT4 SOX2 侵袭转移 

分 类 号：R697+.3[医药卫生—泌尿科学]