中国组织工程研究Issue(28)：5108-5112,5.DOI:10.3969/j.issn.2095-4344.2013.28.002
miR-376b-3p可促进Runx2诱导C2C12细胞进行成骨细胞早期分化*★
miR-376b-3p promotes Runx2-induced early osteogenic differentiation of C2C12 cells
摘要
Abstract
BACKGROUND: The transcription factor Runx2 is the key factor that regulates osteogenic differention and bone development. It has been reported that the C2C12 mesenchymal cells can be induced to differentiate into osteoblasts by Runx2 overexpression, but the molecular mechanism of induction is stil largely unclear. OBJECTIVE: To investigate the role of the members of the miR-376 family during Runx2-induced osteogenic differentiation in C2C12 cells. METHODS: The expression of the members of the miR-376 family was detected by real-time quantitative PCR at different time points using C2C12/Runx2Dox sub-line with conditional Runx2 expression. In miR-376b-3p-transfected C2C12/Runx2Dox cells, the expression of osteoblast markers, such as alkaline phosphatase and osteocalcin, was detected by real-time quantitative PCR, and the alkaline phosphatase activity was also examined by alkaline phosphatase staining. The putative miR-376b-3p targets were commonly predicted by online tools (miRanda, miRWalk and TargetScan). The functional classification of these putative targets was performed by DAVID Bioinformatics Resources database. RESULTS AND CONCLUSION: The expression of miR-376b-3p was significantly increased during Runx2-induced osteogenic differentiation of C2C12 cells, but the expression of other members was not changed. Transfection of miR-376b-3p mimic upregulated alkaline phosphatase expression, but had no effect on osteocalcin expression. The alkaline phosphatase activity was also increased by transfection of miR-376b-3p. The functional classification of miR-376b-3p putative targets showed that miR-376b-3p is involved in the skeleton development, indicating the role of miR-376b-3p in osteoblast differentiation. Taken together, these results suggest that Runx2 promotes early osteogenic differentiation in C2C12 cells by regulating the expression of genes related to osteogenic differentiation through upregulation of miR-376b-3p.关键词
组织构建/骨组织构建/Runx2/C2C12 细胞/miR-376b-3p/成骨分化/强力霉素/国家自然科学基金Key words
tissue construction/bone tissue construction/Runx2/C2C12 cells/miR-376b-3p/osteogenic differentiation/doxycycline/National Natural Science Foundation of China分类
医药卫生引用本文复制引用
耿倩倩,余守和,张越,王红梅,孙奋勇..miR-376b-3p可促进Runx2诱导C2C12细胞进行成骨细胞早期分化*★[J].中国组织工程研究,2013,(28):5108-5112,5.基金项目
the National Natural Science Foundation of China, No.81071524* （）
