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miR-145-5p 通过靶向 E2F 转录因子 3 抑制骨肉瘤细胞增殖。

MicroRNA‑145‑5p inhibits osteosarcoma cell proliferation by targeting E2F transcription factor 3.

机构信息

Department of Orthopedics, Guiyang Maternal and Child Health-Care Hospital, Guiyang, Guizhou 550000, P.R. China.

Department of Pathology, Renmin Hospital of Guizhou, Guiyang, Guizhou 550000, P.R. China.

出版信息

Int J Mol Med. 2020 May;45(5):1317-1326. doi: 10.3892/ijmm.2020.4504. Epub 2020 Feb 17.

DOI:10.3892/ijmm.2020.4504
PMID:32323741
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7138290/
Abstract

Osteosarcoma is a common type of bone tumor that primarily occurs in children and young adults. MicroRNA (miRNA/miR) dysregulation is associated with the progression of osteosarcoma; therefore, the aim of the present study was to investigate the biological functions and molecular mechanisms of miR‑145‑5p in osteosarcoma. The expression of miR‑145‑5p in osteosarcoma tissues and cell lines was quantified using reverse transcription‑quantitative PCR (RT‑qPCR). The effect of miR‑145‑5p on the proliferation of osteosarcoma cells was detected using Cell Counting Kit‑8 and colony formation assays, as well as cell cycle distribution analysis. The effect of miR‑145‑5p on tumor growth was further investigated in vivo using a subcutaneous tumor model in nude mice. The interaction between miR‑145‑5p and E2F transcription factor 3 (E2F3) was determined using bioinformatics analysis, a luciferase assay, RT‑qPCR and western blotting. The results revealed that miR‑145‑5p expression was decreased in osteosarcoma cell lines and tissues compared with the corresponding normal controls. Increased miR‑145‑5p expression inhibited the proliferation and colony formation ability of osteosarcoma cells, and induced G1 phase arrest. Furthermore, mice injected with tumor cells overexpressing miR‑145‑5p exhibited smaller tumors than those in the control group. Further investigation revealed that miR‑145‑5p binds to and decreases the expression of E2F3. In addition, the mRNA levels of E2F3 were negatively associated with miR‑145‑5p in osteosarcoma tissues, and increasing E2F3 expression abrogated the inhibitory effects of miR‑145‑5p on osteosarcoma cells. Collectively, the results obtained in the present study suggest that miR‑145‑5p may suppress the progression of osteosarcoma, and may serve as a useful biomarker for the diagnosis of osteosarcoma, as well as a therapeutic target.

摘要

骨肉瘤是一种常见的骨肿瘤类型,主要发生在儿童和青少年中。microRNA (miRNA/miR) 失调与骨肉瘤的进展有关;因此,本研究旨在探讨 miR-145-5p 在骨肉瘤中的生物学功能和分子机制。采用逆转录-定量 PCR (RT-qPCR) 检测骨肉瘤组织和细胞系中 miR-145-5p 的表达。采用细胞计数试剂盒-8 和集落形成实验以及细胞周期分布分析检测 miR-145-5p 对骨肉瘤细胞增殖的影响。采用裸鼠皮下肿瘤模型进一步研究 miR-145-5p 对肿瘤生长的影响。采用生物信息学分析、荧光素酶报告基因检测、RT-qPCR 和 Western blot 检测 miR-145-5p 与 E2F 转录因子 3 (E2F3) 之间的相互作用。结果显示,与相应的正常对照相比,骨肉瘤细胞系和组织中 miR-145-5p 的表达降低。miR-145-5p 表达增加抑制骨肉瘤细胞的增殖和集落形成能力,并诱导 G1 期阻滞。此外,与对照组相比,注射过表达 miR-145-5p 的肿瘤细胞的小鼠肿瘤较小。进一步研究表明,miR-145-5p 与 E2F3 结合并降低其表达。此外,骨肉瘤组织中 E2F3 的 mRNA 水平与 miR-145-5p 呈负相关,增加 E2F3 表达可消除 miR-145-5p 对骨肉瘤细胞的抑制作用。综上所述,本研究结果表明,miR-145-5p 可能抑制骨肉瘤的进展,可作为骨肉瘤诊断的有用标志物和治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c95a/7138290/0e6c444e0fa5/IJMM-45-05-1317-g06.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c95a/7138290/001c716a1e91/IJMM-45-05-1317-g02.jpg
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