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miR-142-5p 通过靶向 PTEN 促进视网膜母细胞瘤细胞的增殖、迁移和侵袭。

MiR-142-5p promotes retinoblastoma cell proliferation, migration and invasion by targeting PTEN.

机构信息

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.

出版信息

J Biochem. 2021 Oct 11;170(2):195-202. doi: 10.1093/jb/mvaa121.

Abstract

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.

摘要

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。

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