【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。更多还原

【Abstract】 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 prostate cancer cells, and inducing apoptosis.更多还原