Ruan Wen-Dong, Wang Pei, Feng Shiqing, Xue Yuan, Zhang Bin
Department of Orthopedics, Tianjin Medical University General Hospital, Heping District, Tianjin, People's Republic of China.
Onco Targets Ther. 2016 Jan 18;9:303-13. doi: 10.2147/OTT.S95204. eCollection 2016.
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.
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。