The Wnt Signaling Pathway Is Effect On Regulation Of Mouse Osteoclast Differentiation

Objective To make sure whether the Wnt signaling pathway relay onthe role of osteoblasts directly regulates osteoclast differentiation andmaturation or not. Comprehensive understanding of the role of the Wntsignaling pathway in the development of bone state. Lay a solidfoundation for our future use of the Wnt inhibitor antibody treatment ofosteoporosis. On this basis, explicit involvement of the Wnt signalingpathway directly regulates osteoclast differentiation and maturation.Methods Cells were divided into a negative control group (blank), anexperimental control group (Ad-GFP transfection), a positive control group(50ng/mL RANKL induction), and an experimental group （Ad-Wnt3atransfection, Ad-β-catenin transfection, Ad-sost transfection, Ad-Wnt5btransfection, Ad-Wnt7a transfection, Ad-Wnt7b transfection. All the cellswere conventionally cultured for6d. Tartrate-resistant acid phosphatase(TRAP) staining was applied to observe osteoclast differentiation andmaturation. Western Blot analysis was applied to detect the proteinexpression of TRAP and Cathepsin K. Total cellular RNA was extractedafter treatment for3,6and9d, and quantitative real-time PCR was applied to determine the expression of receptor activator of nuclear factorκB(RANK), TRAP, Cathepsin K and matrix metalloproteinase (MMP-9).Results TRAP staining showed that50ng/ml RANKL couldsuccessfully induce RAW264.7cells into multinucheated cells and theAd-sost group cells are also into multinucheated (nuclear≥10). A fewmultinucleated cells (3≤nuclear≤5) were observed in the control group andthe Ad-GFP group, and,mononuclear cells and few dual–nuclear cellswere observed in the Ad-Wnt3a group and in the Ad-β-catenin group.RT-PCR analysis the Ad-wnt3a group and the Ad-β-catenin groupsignificantly lowered RANK, TRAP, Cathepsin K, MMP-9gene expression.Western blot protein detection the TRAP, Cathepsin K the expression wassignificantly reduced. The difference was statistically significant (p﹤0.05).Conclusion Wnt signaling pathway can not be adopted osteoblastsindirectly regulate osteoclast differentitation and maturation. Raw264.7celldifferentiation and maturation withβ-catenin can be suppressed, as same asWnt3a. But the Ad-Spst group can promote the differentiation of mature.