25-hydroxycholesterol promotes RANKL-induced osteoclastogenesis through coordinating NFATc1 and Sp1 complex in the transcription of miR-139-5p
Authors: Yinping Lv
Yanliang Lin
Lishan Zhang
Guozhe Xian
Journal: Biochemical and Biophysical Research Communications (2017)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: 25-hydroxycholesterol; NFATc1; Osteoclast; RANKL; Sp1
Affiliations: Department of Hand and Foot Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China., Department of Medical Engineering, Jinan 250021, China., Department of Hepatobiliary Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China., Center Laboratory, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, China. Electronic address: .

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. 

Retracted: No

PubPeer comments:
 Figure 3D. * Some groups of cells appear to be visible multiple times within or across panels. Boxes of the same color highlight several of these similarities. ![file] Sherloq analysis: ![file] ImageTwin analysis: ![file] , 
 Thank you for reminding us. We have been repeating this part of the experiment, and contact the magazine for correction. Thanks again! , 
 Despite the issues raised in comment #1, the journal decided to issue a Corrigendum, September 2020, found here: https: www.sciencedirect.com science article pii S0006291X2031723X  The authors regret that the Figure 3D were mistakenly uploaded in the original publication. The authors have revised the Figure 3D by placing the correct images. The corresponding Fig 3E has been revised. These corrections do not affect the conclusion in the manuscript. The authors would like to apologize for any inconvenience caused. ,