Luo Xiao-Ji, Tang Da-Gang, Gao Tian-Le, Zhang Yan-Liang, Wang Miao, Quan Zheng-Xue, Chen Jin
Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, P. R. China.
Cell Physiol Biochem. 2014;34(6):2180-8. doi: 10.1159/000369661. Epub 2014 Dec 2.
Multiple MicroRNAs (miRNAs) have been identified in the development and progression of osteosarcoma. However, the expression and roles of miR-212 in osteosarcoma remain largely undefined.
Real-time PCR assays were used to detect the expression of miR-212 in human osteosarcoma tissues. MiR-212 mimics were introduced into MG63 and U2OS cells. Bioinformatic prediction was used to identify the potential targets of miR-212. Protein expression analysis, luciferase assays and rescue assays were used to confirm the substrate of miR-212.
miR-212 was significantly down-regulated in human osteosarcoma tissues, compared with adjacent normal tissues. Introduction of miR-212 mimics into MG63 and U2OS cells inhibited cell proliferation and invasion. Besides, miR-212 overexpression could also inhibit tumor growth in the nude mice. Additionally, bioinformatic prediction suggested that the sex-determining region Y-box 4 (Sox4) is a target gene of miR-212. Sox4 inhibition phenocopied the roles of miR-212, while restored expression of Sox4 dampened miR-212-mediated suppression of tumor progression.
The miR-212/Sox4 interaction plays an important role of in the osteosarcoma progression.
多种微小RNA(miRNA)已在骨肉瘤的发生发展过程中被鉴定出来。然而,miR-212在骨肉瘤中的表达及作用仍不清楚。
采用实时聚合酶链反应(PCR)检测人骨肉瘤组织中miR-212的表达。将miR-212模拟物导入MG63和U2OS细胞。利用生物信息学预测鉴定miR-212的潜在靶标。通过蛋白质表达分析、荧光素酶检测和拯救实验来确认miR-212的作用底物。
与相邻正常组织相比,miR-212在人骨肉瘤组织中显著下调。将miR-212模拟物导入MG63和U2OS细胞可抑制细胞增殖和侵袭。此外,miR-212过表达还可抑制裸鼠体内肿瘤生长。另外,生物信息学预测表明性别决定区Y框蛋白4(Sox4)是miR-212的靶基因。抑制Sox4可模拟miR-212的作用,而恢复Sox4的表达则可减弱miR-212介导的对肿瘤进展的抑制作用。
miR-212/Sox4相互作用在骨肉瘤进展中起重要作用。