Liu Jian, Chen Bohua, Yue Bin, Yang Junde
Department of Orthopedics, Eighth People's Hospital of Qingdao, Medical School of Qingdao University, Qingdao, Shandong 266100, P.R. China.
Exp Ther Med. 2016 Dec;12(6):4135-4141. doi: 10.3892/etm.2016.3880. Epub 2016 Nov 7.
MicroRNAs (miRNAs) are a class of small non-coding RNAs that function as critical gene regulators by targeting the 3' untranslated region (UTR) of mRNA, causing translational repression or mRNA degradation. Deregulation of specific miRNAs, including miR-212, has been identified in patients with osteosarcoma. However, the underlying mechanism is yet to be fully elucidated. The present study aimed to reveal the regulatory mechanism of miR-212 in osteosarcoma cell viability and migration. Quantitative polymerase chain reaction data revealed that miR-212 was significantly downregulated in osteosarcoma tissues compared with normal bone tissues. miR-212 was also downregulated in osteosarcoma cell lines compared with normal osteoblast cell lines. Overexpression of miR-212 significantly suppressed the viability and migration of human osteosarcoma MG-63 and Saos-2 cell lines. In addition, forkhead box protein A1 (FOXA1), an oncogene in osteosarcoma, was predicted to be a putative target of miR-212 by bioinformatical analysis. Furthermore, luciferase reporter assay data confirmed that miR-212 could directly bind to the seed sequences within the 3'UTR of FOXA1 mRNA, and miR-212 negatively mediated the protein levels of FOXA1 in osteosarcoma MG-63 and Saos-2 cells. Moreover, knockdown of FOXA1 also led to a significant decrease in the viability and migration of osteosarcoma MG-63 and Saos-2 cells and the expression levels of FOXA1 were significantly upregulated in osteosarcoma tissues and cell lines. These data suggest that miR-212 inhibits the viability and migration of osteosarcoma cells by targeting FOXA1. Accordingly, miR-212 may become a potential candidate for osteosarcoma therapy.
微小RNA(miRNA)是一类小的非编码RNA,通过靶向mRNA的3'非翻译区(UTR)发挥关键基因调节作用,导致翻译抑制或mRNA降解。在骨肉瘤患者中已发现特定miRNA(包括miR-212)失调。然而,其潜在机制尚未完全阐明。本研究旨在揭示miR-212在骨肉瘤细胞活力和迁移中的调节机制。定量聚合酶链反应数据显示,与正常骨组织相比,miR-212在骨肉瘤组织中显著下调。与正常成骨细胞系相比,miR-212在骨肉瘤细胞系中也下调。miR-212的过表达显著抑制了人骨肉瘤MG-63和Saos-2细胞系的活力和迁移。此外,通过生物信息学分析预测,骨肉瘤中的癌基因叉头框蛋白A1(FOXA1)是miR-212的假定靶标。此外,荧光素酶报告基因检测数据证实,miR-212可直接结合FOXA1 mRNA 3'UTR内的种子序列,并且miR-212负向调节骨肉瘤MG-63和Saos-2细胞中FOXA1的蛋白水平。此外,敲低FOXA1也导致骨肉瘤MG-63和Saos-2细胞的活力和迁移显著降低,并且FOXA1的表达水平在骨肉瘤组织和细胞系中显著上调。这些数据表明,miR-212通过靶向FOXA1抑制骨肉瘤细胞的活力和迁移。因此,miR-212可能成为骨肉瘤治疗的潜在候选物。