The role and mechanism of c-fos in osteoclast differentiation
ZHU Wei;SUN Qin;ZHANG Bo-ran;MA Jing-zhi;LI Wen-qiang;Department of Stomatology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology;
Objective: To investigate the effect of c-fos on the differentiation of RAW264. 7 cells into osteoclasts and its possible epigenetic mechanism.Methods: Murine RAW264. 7 cells were conducted by seeding cells on six-well plates in osteoclast differentiation medium which consisted of 10% FBS,supplemented with 50 ng/m L recombinant mouse-RANKL and30 ng/m L recombinant mouse-M-CSF.Differentiated cells were harvested at designated times for TRAP staining and mRNA extraction.TRAP staining was performed to detect the formation of osteoclasts. Real-time quantitative PCR( qRT-PCR) was used to detect the mRNA expression level of c-fos and osteoclast differentiation-related genes.After overexpression and downregulation of c-fos,RAW264. 7 cells were induced by M-CSF and RANKL. Then TRAP staining and qRT-PCR were performed to detect the formation of osteoclasts and the expression level of osteoclast differentiation-related genes. DIANA,miRDB,PicTar and Target Scan databases were conducted to predict microRNAs that may regulate c-fos expression.Results:c-fos was gradually up-regulated during osteoclast differentiation. Overexpression of c-fos promoted the differentiation of RAW264. 7 cells into osteoclasts,while inhibition of c-fos did the opposite effect.Bioinformatics analysis suggested that miR-101a-3p and miR-101b-3p might play a pivotal role in the regulation of c-fos gene expression.Conclusion: c-fos promoted the differentiation of RAW264. 7 cells,and c-fos might be regulated by both miR-101a-3p and miR-101b-3p during this process.