关键词: 关键词：Wnt/β-catenin，RAW264.7，RANKL ，TRAP

Abstract: Objective To learn the regulation of Wnt/β-catenin signaling pathway in the mouse monocyte/macrophage Raw264.7 differentiation and maturation. Methods The cells were divided into four groups: the negative control group (empty Raw264.7), the experimental control group (infected with Ad-GFP), the positive control group (50ng/mL RANKL induced) and the experimental group (infected with Ad-β-catenin). RNA were extracted after the cells were treated for 3, 6, 9days respectively, The expressions of nuclear factor KB receptor, Tartrate-resistant acid phosphatase, Cathepsin K and Metalloproteinase-9 gene were detected by fluorescence quantitative RT-PCR respectively. Cells treated for 6 days were used to observe the differentiation of Raw264.7 by Tartaric acid-resistant phosphatase (TRAP) staining and expressions of TRAP and Cathepsin K protein by Western blot. Results qRT-PCR showed that Ad-β-catenin group could decrease the mRNA expression of RANK, TRAP, Cathepsin K and MMP-9 gene. TRAP staining showed that 50ng/mL RANKL could successfully induced Raw264.7 into multi-cores. Few multinucleated cells in the control group and the Ad-GFP group were seen, In Ad-β-catenin group cells were mononuclear. The TRAP and Cathepsin K protein expression was significantly reduced analysed by Western blot ( p<0.05). Conclusion Wnt/β-catenin signaling pathway can inhibit Raw264.7 differentiate into mature osteoclasts.

Key words: Keyword: Wnt/β-catenin RAW264.7 RANKL TRAP