ABSTRACT
PURPOSE: Based on the Cre-Loxp gene knockout system, this study intended to construct tamoxifen-inducible STAT3 conditional knockout mice and verify the knockout efficiency. METHODS: The inducible osteoblasts-specific Stat3 knockout mice Stat3Col1ERT2 were obtained by hybridization through C57 mice of Stat3fl/fl and Col1 creERT2. Bone mesenchymal stem cells(BMSCs) of these mice were isolated and cultured with or without 4-hydroxytamoxin(4-OTH), to verify the effect of Stat3 knockout in vitro by real-time quantitative PCR and Western blotting in the level of mRNA and protein. Meanwhile, wild type and Stat3Col1ERT2 mice were both intraperitoneally injected with tamoxifen, the expression of STAT3 in the maxillary alveolar bone was observed by immunofluorescent staining to confirm the knockout effect in vivo. Statistical analysis was conducted with SPSS 24.0 software package. RESULTS: Real-time quantitative PCR and Western blotting results demonstrated that mRNA(P<0.05) and protein levels of STAT3 were significantly decreased (P<0.05) in BMSCs derived from Stat3Col1ERT2 mice by 4-OHT induced knockout in vitro. Immunofluorescent staining indicated that STAT3 expression was significantly reduced(P<0.05) in osteoblasts of the maxillary alveolar bone in Stat3Col1ERT2 mice. CONCLUSIONS: This study successfully constructed the inducible osteoblasts-specific Stat3 gene knockout mice, which helped investigators control the time and space of gene knockout, therefore providing new insights and guidance for research fields of orthodontic tooth movement, distraction osteogenesis and jaw fractures in the future.
