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MicroRNA-744 通过靶向 PTEN 促进骨肉瘤细胞的增殖。

MicroRNA-744 promotes proliferation of osteosarcoma cells by targeting PTEN.

机构信息

Department of Orthopedic Surgery, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.

Department of Orthopedic Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, P.R. China.

出版信息

Mol Med Rep. 2020 May;21(5):2276-2282. doi: 10.3892/mmr.2020.11030. Epub 2020 Mar 18.

Abstract

MicroRNAs (miRNAs/miRs) are non-coding RNAs that regulate protein synthesis by targeting mRNAs for translational repression or degradation. Previous studies have reported that aberrant expression of miR‑744 may be involved in human osteosarcoma; however, the underlying mechanisms remain elusive. In the present study, the expression levels of miR‑744 and its downstream signals were determined by reverse transcription‑quantitative PCR and western blotting. Cell proliferation was assessed using the bromodeoxyuridine assay, and the target of miR‑744 was investigated using a dual‑luciferase activity assay. The present study identified a significant upregulation of miR‑744 in osteosarcoma tissues compared with adjacent non‑tumor tissues. Furthermore, it was demonstrated that ectopic overexpression of miR‑744 induced by a miR‑744 precursor significantly enhanced proliferation of the osteosarcoma cell line MG63, whereas opposite results were observed following suppression of miR‑744 with its inhibitor. Moreover, as a unique anti‑oncogene, PTEN was identified as a direct target of miR‑744. It was confirmed that miR‑744 downregulated PTEN expression in MG63 cells by targeting the PTEN 3'untranslated region, and that the downstream AKT signal was also regulated by miR‑744. Collectively, the present results suggested that miR‑744 promoted proliferation of human osteosarcoma cells by directly regulating the PTEN/AKT signaling pathway.

摘要

微小 RNA(miRNAs/miRs)是一类非编码 RNA,通过靶向 mRNAs 来抑制翻译或降解,从而调节蛋白质合成。先前的研究报道,miR-744 的异常表达可能与人类骨肉瘤有关,但其潜在机制尚不清楚。在本研究中,通过逆转录-定量 PCR 和 Western blot 测定了 miR-744 的表达水平及其下游信号。使用溴脱氧尿苷测定法评估细胞增殖,并用双荧光素酶活性测定法研究 miR-744 的靶标。本研究发现骨肉瘤组织中 miR-744 的表达水平明显上调,与相邻非肿瘤组织相比。此外,研究表明,miR-744 前体过表达 miR-744 可显著增强骨肉瘤细胞系 MG63 的增殖,而抑制 miR-744 则观察到相反的结果。此外,作为一种独特的抑癌基因,PTEN 被鉴定为 miR-744 的直接靶标。研究证实,miR-744 通过靶向 PTEN 3'非翻译区下调 MG63 细胞中的 PTEN 表达,并且 miR-744 还调节下游 AKT 信号。综上所述,本研究结果表明,miR-744 通过直接调节 PTEN/AKT 信号通路促进人类骨肉瘤细胞的增殖。

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