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miR-486-3p 通过靶向 ECM1 抑制视网膜母细胞瘤细胞的增殖、迁移和侵袭。

MiR-486-3p inhibits the proliferation, migration and invasion of retinoblastoma cells by targeting ECM1.

机构信息

Department of Ophthalmology, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China.

School of Mechanical Engineering and Automation, Northeastern University, Shenyang, Liaoning, China.

出版信息

Biosci Rep. 2020 Jun 26;40(6). doi: 10.1042/BSR20200392.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a580/7273916/fe399cfc6118/bsr-40-bsr20200392-g1.jpg

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