Department of Ophthalmology, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China.
School of Mechanical Engineering and Automation, Northeastern University, Shenyang, Liaoning, China.
Biosci Rep. 2020 Jun 26;40(6). doi: 10.1042/BSR20200392.
It has been reported that miR-486-3p expression is decreased in retinoblastoma (RB) tumor tissues, however, its function in RB has been less reported. The present study aimed to explore the regulatory effects of miR-486-3p on RB cells. The expression of miR-486-3p in RB tissues and cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability, proliferation, apoptosis, migration and invasion ability were determined by cell counting kit-8 (CCK-8) kit, clone formation assay, flow cytometry, scratch assay and transwell, respectively. Targetscan 7.2 and dual-luciferase reporter were used to verify target genes for miR-486-3p. The expressions of apoptosis-related proteins and ECM1 were detected by Western blot. The miR-486-3p expression was decreased in RB tissues and cells. In RB cells, overexpression of miR-486-3p inhibited cell proliferation, migration and invasion, while promoted apoptosis. Moreover, overexpression of miR-486-3p decreased Bcl-2 expression, while increased the expressions of Bax and Cleaved Caspase-3 (C caspase-3). ECM1 was the target gene of miR-486-3p, and miR-486-3p inhibited the expression of ECM1. Furthermore, ECM1 partially reversed the inhibitory effect of miR-486-3p on the proliferation, migration and invasion of RB cells. MiR-486-3p inhibited the proliferation, migration and invasion of RB by down-regulating ECM1.
据报道,miR-486-3p 在视网膜母细胞瘤 (RB) 肿瘤组织中的表达降低,但其在 RB 中的功能报道较少。本研究旨在探讨 miR-486-3p 对 RB 细胞的调控作用。采用实时定量聚合酶链反应 (qRT-PCR) 检测 miR-486-3p 在 RB 组织和细胞中的表达。通过细胞计数试剂盒 (CCK-8) 试剂盒、克隆形成实验、流式细胞术、划痕实验和 Transwell 分别测定细胞活力、增殖、凋亡、迁移和侵袭能力。Targetscan 7.2 和双荧光素酶报告基因用于验证 miR-486-3p 的靶基因。Western blot 检测凋亡相关蛋白和 ECM1 的表达。miR-486-3p 在 RB 组织和细胞中表达降低。在 RB 细胞中,过表达 miR-486-3p 抑制细胞增殖、迁移和侵袭,促进细胞凋亡。此外,过表达 miR-486-3p 降低 Bcl-2 表达,增加 Bax 和 Cleaved Caspase-3 (C caspase-3) 的表达。ECM1 是 miR-486-3p 的靶基因,miR-486-3p 抑制 ECM1 的表达。此外,ECM1 部分逆转了 miR-486-3p 对 RB 细胞增殖、迁移和侵袭的抑制作用。miR-486-3p 通过下调 ECM1 抑制 RB 的增殖、迁移和侵袭。