Inhibition of osteoclastogenesis by RNA interference targeting RANK

BMC Musculoskeletal Disorders

Christian Wedemeyer³  Marcus Jäger³  Max Daniel Kauther¹  Bin Dong²  Jie Xu²  Ruofan Ma² 

[1] Department of Trauma Surgery, University of Duisburg-Essen, Hufelandstr. 55, 45122, Essen, Germany;Department of Orthopaedics, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 West Yan Jiang Road, PO Box 510120, Guangzhou, China;Department of Orthopaedics, University of Duisburg-Essen, Hufelandstr. 55, 45122, Essen, Germany

关键词: Cell culture;    RNA interference;    Inhibition;    RANK;   

Others  :  1145925
DOI  :  10.1186/1471-2474-13-154

 received in 2011-11-25, accepted in 2012-08-21,  发布年份 2012

【 摘 要 】

Background

Osteoclasts and osteoblasts regulate bone resorption and formation to allow bone remodeling and homeostasis. The balance between bone resorption and formation is disturbed by abnormal recruitment of osteoclasts. Osteoclast differentiation is dependent on the receptor activator of nuclear factor NF-kappa B (RANK) ligand (RANKL) as well as the macrophage colony-stimulating factor (M-CSF). The RANKL/RANK system and RANK signaling induce osteoclast formation mediated by various cytokines. The RANK/RANKL pathway has been primarily implicated in metabolic, degenerative and neoplastic bone disorders or osteolysis. The central role of RANK/RANKL interaction in osteoclastogenesis makes RANK an attractive target for potential therapies in treatment of osteolysis. The purpose of this study was to assess the effect of inhibition of RANK expression in mouse bone marrow macrophages on osteoclast differentiation and bone resorption.

Methods

Three pairs of short hairpin RNAs (shRNA) targeting RANK were designed and synthesized. The optimal shRNA was selected among three pairs of shRNAs by RANK expression analyzed by Western blot and Real-time PCR. We investigated suppression of osteoclastogenesis of mouse bone marrow macrophages (BMMs) using the optimal shRNA by targeting RANK.

Results

Among the three shRANKs examined, shRANK-3 significantly suppressed [88.3%] the RANK expression (p < 0.01). shRANK-3 also brought about a marked inhibition of osteoclast formation and bone resorption as demonstrated by tartrate–resistant acid phosphatase (TRAP) staining and osteoclast resorption assay. The results of our study show that retrovirus-mediated shRANK-3 suppresses osteoclast differentiation and osteolysis of BMMs.

Conclusions

These findings suggest that retrovirus-mediated shRNA targeting RANK inhibits osteoclast differentiation and osteolysis. It may appear an attractive target for preventing osteolysis in humans with a potential clinical application.

【 授权许可】

   
2012 Ma et al.; licensee BioMed Central Ltd.

【 图 表 】

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