miRNA-150-5p 通过下调 VEGFA 抑制骨肉瘤细胞的增殖和侵袭。

MicroRNA-150-5p inhibits proliferation and invasion of osteosarcoma cells by down-regulating VEGFA.

机构信息

Department of Orthopedics, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9265-9273. doi: 10.26355/eurrev_202009_23008.

DOI:10.26355/eurrev_202009_23008

PMID:33015767

Abstract

OBJECTIVE

The aim of this study was to investigate whether microRNA-150-5p was involved in osteosarcoma cell proliferation and invasiveness via modulating vascular endothelial growth factor A (VEGFA) expression.

PATIENTS AND METHODS

10 pairs of osteosarcoma tissues and para-cancerous tissues were collected from patients with osteosarcoma in our center from February 2012 to July 2018. Relative expression levels of microRNA-150-5p and VEGFA in tissues and cell lines were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Luciferase reporter gene assay was conducted to illustrate the binding interplay between microRNA-150-5p and VEGFA. Furthermore, proliferative and invasive potentials in HOS and MG-63 cells regulated by both microRNA-150-5p and VEGFA were determined using Cell Counting Kit-8 (CCK-8), colony formation assay, and transwell assay, respectively.

RESULTS

MicroRNA-150-5p was remarkably downregulated, while VEGFA was upregulated in osteosarcoma tissues compared with para-cancerous tissues (p<0.05). Similar results were observed in osteosarcoma cells and normal osteoblasts. Overexpression of microRNA-150-5p significantly inhibited the proliferation and invasion of osteosarcoma cells (p<0.05). Luciferase reporter gene assay demonstrated that microRNA-150-5p could target to VEGFA to negatively modulate its expression. In addition, the knockdown of VEGFA remarkably weakened osteosarcoma cell proliferative and invasive capacities (p<0.05).

CONCLUSIONS

MicroRNA-150-5p weakens proliferative and invasive potentials in osteosarcoma cells by downregulating VEGFA level. All our findings suggest that microRNA-150-5p/VEGFA axis is a promising target for osteosarcoma treatment.

摘要

目的

本研究旨在探讨微小 RNA-150-5p 是否通过调节血管内皮生长因子 A（VEGFA）的表达来参与骨肉瘤细胞的增殖和侵袭。

方法

收集 2012 年 2 月至 2018 年 7 月我院收治的 10 对骨肉瘤组织及其癌旁组织。采用实时定量聚合酶链反应（qRT-PCR）检测组织和细胞系中微小 RNA-150-5p 和 VEGFA 的相对表达水平。通过荧光素酶报告基因实验阐明微小 RNA-150-5p 与 VEGFA 之间的结合相互作用。此外，通过细胞计数试剂盒-8（CCK-8）、集落形成实验和 Transwell 实验分别确定微小 RNA-150-5p 和 VEGFA 对 HOS 和 MG-63 细胞增殖和侵袭能力的调节作用。

结果

与癌旁组织相比，骨肉瘤组织中微小 RNA-150-5p 显著下调，而 VEGFA 上调（p<0.05）。在骨肉瘤细胞和正常成骨细胞中也观察到了类似的结果。过表达微小 RNA-150-5p 可显著抑制骨肉瘤细胞的增殖和侵袭（p<0.05）。荧光素酶报告基因实验表明，微小 RNA-150-5p 可以靶向 VEGFA 以负调控其表达。此外，VEGFA 的敲低显著减弱了骨肉瘤细胞的增殖和侵袭能力（p<0.05）。