Ye Jing, Xie Wei, Zuo Yunzhou, Jing Guangwu, Tong Jie
Department of Orthopedics, Hubei 672 Orthopedics Hospital of Integrated Chinese and Western Medicine, Wuhan, Hubei 430079, P.R. China.
Department of Orthopedics, The Sixth Hospital of Wuhan Affiliated to Jianghan University, Wuhan, Hubei 430015, P.R. China.
Exp Ther Med. 2020 Feb;19(2):1425-1431. doi: 10.3892/etm.2019.8356. Epub 2019 Dec 19.
MicroRNAs (miRNAs) are integrally involved in biological and pathobiological development. Many studies have demonstrated the abnormal expression of microRNA-496 (miR-496) in various human malignant tumors. The present study was designed to investigate the functions and the underlying mechanisms of miR-496 in osteosarcoma (OS) progression. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to determine the expression of miR-496 in OS tissues and cell lines. Luciferase activity was used to confirm the interaction between miR-496 and brain derived neurotrophic factor (), a downstream gene of miR-496. RT-qPCR was also used to quantify mRNA expression, and the BDNF protein expression level was detected by western blot analysis. In addition, the Cell Counting Kit-8 (CCK-8) was used to detect cell viability. The results revealed that the level of miR-496 expression was significantly reduced in osteosarcoma tissues and cell lines. was verified to be a direct target gene of miR-496 and was found to be negatively regulated by miR-496. Overall, it was demonstrated that miR-496 inhibits osteosarcoma cell proliferation via inhibition of BDNF. Thus, the miR-496/BDNF axis may be a novel strategy for the clinical treatment of OS.
微小RNA(miRNA)全面参与生物和病理生物学发展。许多研究已证明微小RNA-496(miR-496)在各种人类恶性肿瘤中表达异常。本研究旨在探讨miR-496在骨肉瘤(OS)进展中的功能及潜在机制。采用逆转录-定量聚合酶链反应(RT-qPCR)检测miR-496在OS组织和细胞系中的表达。利用荧光素酶活性证实miR-496与脑源性神经营养因子(BDNF)(miR-496的下游基因)之间的相互作用。RT-qPCR还用于定量BDNF mRNA表达,通过蛋白质印迹分析检测BDNF蛋白表达水平。此外,使用细胞计数试剂盒-8(CCK-8)检测细胞活力。结果显示,骨肉瘤组织和细胞系中miR-496表达水平显著降低。BDNF被证实为miR-496的直接靶基因,且发现其受miR-496负调控。总体而言,证明miR-496通过抑制BDNF抑制骨肉瘤细胞增殖。因此,miR-496/BDNF轴可能是OS临床治疗的新策略。