title: 25-hydroxycholesterol promotes RANKL-induced osteoclastogenesis through coordinating NFATc1 and Sp1 complex in the transcription of miR-139-5p.
abstract: 25-hydroxycholesterol (25-HC) is implicated in many processes, including lipid metabolism and the immune response. However, the role of 25-HC in RANKL-induced osteoclastogenesis remains largely unknown. Our results showed that 25-HC inhibited miR-139-5p expression in mouse bone marrow macrophages (BMMs) cultured in receptor activator of NF-κB ligand (RANKL) and monocyte macrophage colony-stimulating factor (M-CSF). Further investigation suggested that 25-HC promoted the expression of nuclear factor of activated T cell cytoplasmic 1 (NFATc1) and Sp1, especially in the presence of RANKL and M-CSF. Meanwhile, 25-HC induced nuclear translocation of NFATc1, resulting in the interaction between NFATc1 and Sp1 that was confirmed by co-immunoprecipitation. Chromatin immunoprecipitation assay indicated that Sp1 could bind to miR-139-5p promoter, but NFATc1 had no binding capacity. Although forming NFATc1/Sp1 complex increased its binding to miR-139-5p promoter, the complex inhibited the transcriptional activity of Sp1. Inhibition of NFATc1 increase the expression of miR-139-5p, which might be due to the release of free Sp1 that could bind to the promoter of miR-139-5p. Enforced expression of miR-139-5p impaired osteoclastogenesis induced by co-treatment with 25-HC and RANKL. These results suggested that 25-HC induced the interaction between NFATc1 and Sp1, reducing the level of free Sp1 to inhibit miR-139-5p expression and promote osteoclastogenesis.
authors.affiliation: Department of Hand and Foot Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China.
authors.forename: Lishan
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Medical Engineering, Jinan 250021, China.
authors.forename: Yinping
authors.initials: Y
authors.lastname: Lv
authors.affiliation: Department of Hepatobiliary Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China.
authors.forename: Guozhe
authors.initials: G
authors.lastname: Xian
authors.affiliation: Center Laboratory, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China. Electronic address: sphyanlianglin@163.com.
authors.forename: Yanliang
authors.initials: Y
authors.lastname: Lin
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D006888:Hydroxycholesterols / Q000494:pharmacology*; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics*; D018613:Microscopy, Confocal; D050778:NFATC Transcription Factors / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010010:Osteoclasts / Q000187:drug effects* / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D011485:Protein Binding; D053245:RANK Ligand / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016329:Sp1 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D006888:Hydroxycholesterols; C546202:MIRN139 microRNA, mouse; D035683:MicroRNAs; D050778:NFATC Transcription Factors; C494005:Nfatc1 protein, mouse; D053245:RANK Ligand; D016329:Sp1 Transcription Factor; C007997:25-hydroxycholesterol; D016173:Macrophage Colony-Stimulating Factor
keywords: 25-hydroxycholesterol; NFATc1; Osteoclast; RANKL; Sp1; miR-139-5p