lncRNASOX2-OT调节miR-215-5p/ZEB2轴抑制人前列腺癌细胞系增殖和上皮间质转化研究  

The study of long non-coding RNA SRY-box transcription factor 2 overlapping transcript regulates the miR-215-5p/zinc finger E-box-binding protein 2 axis to suppress proliferation and epithelial-mesenchymal transition in human prostate cancer cell lines

作　　者：史春辉 李波[1] 朱星宇[2] 何仲琴 SHI Chunhui;LIBo;ZHU Xingyu;HE Zhongqin(Department of Oncology,Baoji Hospital of Traditional Chinese Medicine,Baoji 721000,Shaanxi,China;Department of Respiratory Medicine,the Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,Shaanxi,China)

机构地区：[1]宝鸡市中医医院肿瘤科,陕西宝鸡721000 [2]陕西中医药大学第二附属医院呼吸内科,陕西咸阳712000

出　　处：《中国性科学》2023年第12期24-32,共9页Chinese Journal of Human Sexuality

基　　金：宝鸡市卫生和计划生育项目(2016-28)。

摘　　要：目的探讨长非编码RNA SRY盒转录因子2重叠转录本(lncRNA SOX2-OT)调节miR-215-5p/E盒结合锌指蛋白2(ZEB2)轴对前列腺癌细胞增殖和上皮间质转化(EMT)的影响。方法将PC-3细胞分为对照组、si-SOX2-OT组、si-NC组、si-SOX2-OT+anti-miR-215-5p组,实时荧光定量聚合酶链反应(qRT-PCR)检测细胞中SOX2-OT、miR-215-5p与ZEB2表达。细胞计数试剂盒-8(CCK-8)及5-乙炔基-2′脱氧尿嘧啶核苷(EdU)染色法检测PC-3细胞增殖;流式细胞术检测PC-3细胞凋亡率;Transwell检测PC-3细胞迁移、侵袭;qRT-PCR检测PC-3细胞miR-215-5p、ZEB2 mRNA、凋亡蛋白与EMT标志蛋白转录表达;Western blot检测PC-3细胞ZEB2、凋亡蛋白与EMT标志蛋白表达;双荧光素酶报告基因实验验证PC-3细胞中SOX2-OT对miR-215-5p及miR-215-5p对ZEB2的靶向调节作用。结果与人前列腺上皮细胞相比,人前列腺癌组织源细胞和人前列腺癌细胞株PC-3、DU 145、22RV1中SOX2-OT、ZEB2 mRNA表达升高,miR-215-5p表达降低(P<0.05)。与对照组相比,si-SOX2-OT组细胞活力、EdU阳性率、迁移细胞数、侵袭细胞数、细胞ZEB2、B淋巴细胞瘤-2基因(Bcl-2)及Vimentin mRNA和蛋白表达均降低,细胞凋亡率、miR-215-5p及Bcl-2相关X基因(Bax)、E-cadherin mRNA和蛋白表达均升高(P<0.05)。下调miR-215-5p表达可减弱敲低SOX2-OT对PC-3细胞的影响。结论敲低SOX2-OT可通过促进miR-215-5p表达而下调ZEB2的表达,从而抑制前列腺癌细胞增殖、迁移、侵袭和EMT,并诱导细胞凋亡。Objective To investigate the influences of long non-coding RNA SRY-box transcription factor 2 overlapping transcript(lncRNA SOX2-OT)on the proliferation and epithelial-mesenchymal transition(EMT)of prostate cancer cells by regulating miR-215-5p/zinc finger E-box-binding protein 2(ZEB2)axis.Methods PC-3 cell was divided into control group,si-SOX2-OT group,si-NC group,si-SOX2-OT+anti-miR-215-5p group.The expression of SOX2-OT,miR-215-5p and ZEB2 in cells was detected by quantitative real time polymerase chain reaction(qRT-PCR).PC-3 cell proliferation was detected by cell counting kit-8(CCK-8)and 5-Ethynyl-2′-deoxyuridine(EdU)staining;the apoptosis rate of PC-3 cells was detected by flow cytometry;the migration and invasion of PC-3 cells were detected by Transwell;the transcription and expression of miR-215-5p,ZEB2 mRNA,apoptosis protein and EMT marker protein in PC-3 cells were detected by qRT-PCR;the expressions of ZEB2,apoptotic protein and EMT marker protein in PC-3 cells were detected by Western blot;double luciferase reporter gene experiment verified the targeted regulation of SOX2-OT on miR-215-5p and miR-215-5p on ZEB2 in PC-3 cells.Results Compared with human prostate epithelial cells,the expression of SOX2-OT and ZEB2 mRNA in human prostate cancer tissue-derived cells and human prostate cancer cell lines PC-3,DU 145 and 22RV1 were increased,and the expression of miR-215-5p was decreased(P<0.05).Compared with the control group,the cell viability,EdU positive rate,the number of migrating cells,the number of invasive cells,and the mRNA and protrin expression of ZEB2,Bcl-2 and Vimentin in the si-SOX2-OT group were decreased,the apoptosis rate,the expression of miR-215-5p,Bax and E-cadherin mRNA and protrin were increased(P<0.05).Downregulating the expression of miR-215-5p attenuates the effect of SOX2-OT knockdown on PC-3 cells.Conclusions Knockdown of SOX2-OT can downregulate the expression of ZEB2 by promoting the expression of miR-215-5p,thereby inhibiting the proliferation,migration,invasion and EMT of p

关 键 词：长非编码RNA SRY盒转录因子2重叠转录本 miR-215-5p/E盒结合锌指蛋白2轴 前列腺癌 增殖 

分 类 号：R697[医药卫生—泌尿科学]