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微小RNA-497通过靶向人骨肉瘤细胞中的AMOT抑制细胞增殖、迁移和侵袭。

MicroRNA-497 inhibits cell proliferation, migration, and invasion by targeting AMOT in human osteosarcoma cells.

作者信息

Ruan Wen-Dong, Wang Pei, Feng Shiqing, Xue Yuan, Zhang Bin

机构信息

Department of Orthopedics, Tianjin Medical University General Hospital, Heping District, Tianjin, People's Republic of China.

出版信息

Onco Targets Ther. 2016 Jan 18;9:303-13. doi: 10.2147/OTT.S95204. eCollection 2016.

Abstract

MicroRNAs (miRNAs) have a role in the development and progression of human malignancy. The expression of miR-497 is decreased in malignant tumors, which suggests a role for miR-497 as a tumor suppressor. Angiomotin is encoded by the AMOT gene, which is a target for miR-497. Angiomotin has a role in angiogenesis, cell proliferation, and invasion in human malignancies, including osteosarcoma. However, the role of miR-497 in human osteosarcoma is unknown. This preliminary study included human osteosarcoma tissues and normal tissues from 20 patients, the osteosarcoma cell lines, MG-63, SAOS-2, U-2 OS, and the human osteoblast cell line hFOB (OB3). Western blots for angiomotin and quantitative real-time polymerase chain reaction for the expression of miR-497 and AMOT were performed. Knockdown studies were performed using RNA interference and transfection studies used miR-497 mimics. Quantitative cell migration assays were performed, and cell apoptosis was studied by flow cytometry. Osteosarcoma cells and cell lines showed reduced expression of miR-497 and increased expression of angiomotin. Transfection of osteosarcoma cells with miR-497 mimics suppressed the expression of angiomotin. Results from a dual-luciferase reporter system supported AMOT as a direct target gene of miR-497. Knockdown of AMOT using RNA interference resulted in inhibition of osteosarcoma cell proliferation, migration, and invasion. These preliminary studies support a role for miR-497 as a suppressor of AMOT gene expression in human osteosarcoma cells, resulting in suppression of tumor cell proliferation and invasion. Further studies are recommended to investigate the role of miR-497 in osteosarcoma and other malignant mesenchymal tumors.

摘要

微小RNA(miRNA)在人类恶性肿瘤的发生和发展中发挥作用。miR-497在恶性肿瘤中的表达降低,这表明miR-497具有肿瘤抑制作用。血管动蛋白由AMOT基因编码,该基因是miR-497的一个靶标。血管动蛋白在包括骨肉瘤在内的人类恶性肿瘤的血管生成、细胞增殖和侵袭中发挥作用。然而,miR-497在人类骨肉瘤中的作用尚不清楚。这项初步研究纳入了20例患者的人类骨肉瘤组织和正常组织、骨肉瘤细胞系MG-63、SAOS-2、U-2 OS以及人类成骨细胞系hFOB(OB3)。进行了血管动蛋白的蛋白质免疫印迹分析以及miR-497和AMOT表达的定量实时聚合酶链反应。使用RNA干扰进行敲低研究,使用miR-497模拟物进行转染研究。进行了定量细胞迁移试验,并通过流式细胞术研究细胞凋亡。骨肉瘤细胞和细胞系显示miR-497表达降低,血管动蛋白表达增加。用miR-497模拟物转染骨肉瘤细胞可抑制血管动蛋白的表达。双荧光素酶报告系统的结果支持AMOT是miR-497的直接靶基因。使用RNA干扰敲低AMOT可导致骨肉瘤细胞增殖、迁移和侵袭受到抑制。这些初步研究支持miR-497在人类骨肉瘤细胞中作为AMOT基因表达抑制剂的作用,从而导致肿瘤细胞增殖和侵袭受到抑制。建议进一步研究以探讨miR-497在骨肉瘤和其他恶性间叶肿瘤中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3767/4727508/1e875dc418e4/ott-9-303Fig1.jpg

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