Department of Ophthalmology, Affiliated Hospital of Jining Medical University, Jining 272029, China.
Department of Ophthalmology, Shanxian Central Hospital, Wenhua Road No. 1, Heze 274300, Shandong, China.
J Biochem. 2021 Oct 11;170(2):195-202. doi: 10.1093/jb/mvaa121.
The study intends to probe the functions of miR-142-5p in retinoblastoma (RB) and the relationship between miR-142-5p and phosphatase and tensin homolog deleted on chromosome ten (PTEN). In our study, miR-142-5p and PTEN mRNA expression in RB tissue, serum of RB patients and RB cell lines were investigated by quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation, migration, invasion and cell apoptosis were measured using MTT assay, BrdU assay, Transwell experiments and flow cytometry analysis, respectively. Binding sites between miR-142-5p and PTEN were predicted by the TargetScan database and were confirmed via qRT-PCR, western blot and dual-luciferase reporter gene assay. It was demonstrated that miR-142-5p expression was elevated in RB tissue, serum of RB patients and RB cell lines. MiR-142-5p overexpression remarkably promoted the proliferation, migration, invasion and inhibited the apoptosis of WERI-RB-1 cells while miR-142-5p knockdown induced opposite effects in Y79 cells. MiR-142-5p decreased PTEN expression in both mRNA and protein expression levels, and PTEN was identified as a target gene of miR-142-5p. Cotransfection of PTEN overexpression plasmids reversed the influences of miR-142-5p on RB cells. In conclusion, miR-142-5p enhances proliferation, migration and invasion of RB cell by targeting PTEN.
本研究旨在探究 miR-142-5p 在视网膜母细胞瘤(RB)中的作用及其与磷酸酶张力蛋白同源物缺失的染色体 ten(PTEN)之间的关系。在我们的研究中,通过定量实时聚合酶链反应(qRT-PCR)检测了 RB 组织、RB 患者血清和 RB 细胞系中 miR-142-5p 和 PTEN mRNA 的表达。通过 MTT 测定、BrdU 测定、Transwell 实验和流式细胞术分析分别测量了增殖、迁移、侵袭和细胞凋亡。miR-142-5p 和 PTEN 之间的结合位点通过 TargetScan 数据库进行预测,并通过 qRT-PCR、western blot 和双荧光素酶报告基因测定进行验证。结果表明,miR-142-5p 在 RB 组织、RB 患者血清和 RB 细胞系中表达升高。miR-142-5p 过表达显著促进了 WERI-RB-1 细胞的增殖、迁移和侵袭,而抑制了 Y79 细胞的凋亡;而 miR-142-5p 下调则诱导了相反的效应。miR-142-5p 降低了 PTEN 在 mRNA 和蛋白表达水平上的表达,并且 PTEN 被鉴定为 miR-142-5p 的靶基因。PTEN 过表达质粒的共转染逆转了 miR-142-5p 对 RB 细胞的影响。综上所述,miR-142-5p 通过靶向 PTEN 增强了 RB 细胞的增殖、迁移和侵袭。