Mutant hFGF23(A12D) stimulates osteoblast differentiation through FGFR3
Mutant hFGF23(A12D) stimulates osteoblast differentiation through FGFR3
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突变体 hFGF23(A12D) 通过 FGFR3 刺激成骨细胞分化
DOI：
10.1111/jcmm.14201
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发表时间：
2019-04-01
影响因子：
5.3
通讯作者：
Chen, Fengshan
中科院分区：
文献类型：
--
作者：
Tu, Yilin;Qu, Taoran;Chen, Fengshan
成纤维细胞生长因子（FGF）23是FGF家族的成员，通过与FGFR相互作用参与骨发育。在以前的研究中，我们发现了一个突变的人成纤维细胞生长因子（hFGF）23（A12 D）的下颌骨无颌症（MP）的家系。然而，hFGF 23（A12 D）在骨形成过程中的确切作用仍不清楚。本研究的目的是鉴定hFGF 23（A12 D）在骨形成中的功能。我们分别用含突变型hFGF 23（A12 D）和野生型hFGF 23的重组慢病毒感染分离的大鼠颅骨（RC）细胞。实时PCR、蛋白质印迹和酶联免疫吸附试验证实hFGF 23（A12 D）未能分泌。通过CCK-8法检测细胞生长情况，碱性磷酸酶染色法检测细胞分化能力，RT-PCR检测hFGF 23（A12 D）对RC细胞增殖有抑制作用，并能促进RC细胞向成骨细胞分化。通过RNA测序、RT-PCR和western blot检测发现FGFR 3的表达增加。通过免疫共沉淀试验和免疫荧光染色，我们发现hFGF 23（A12 D）通过与FGFR 3的胞内结构域相互作用激活丝裂原活化蛋白激酶信号通路。总之，我们确定了hFGF 23（A12 D）参与成骨细胞生成和形成的机制，这特别是由于其与FGFR 3的相互作用。
Fibroblast growth factor (FGF) 23 is a member of the FGF family involved in bone development by interacting with FGFRs. In a previous study, we discovered a mutant human FGF (hFGF) 23 (A12D) in the mandibular prognathism (MP) pedigree. However, the exact role of hFGF23(A12D) during bone formation remains unclear. The aim of this study was to identify the function of hFGF23(A12D) in bone formation. We infected isolated rat calvaria (RC) cells with the recombinant lentivirus containing mutant hFGF23(A12D) and WT hFGF23 respectively. Real-time PCR, western blot and enzyme-linked immunosorbent assay confirmed that hFGF23(A12D) failed to be secreted. We measured cell growth via the CCK-8 assay based on Zsgreen expression, detected cell differentiation ability via alkaline phosphatase staining, performed RT-PCR and found that hFGF23(A12D) inhibited proliferation of RC cells and stimulated the differentiation of RC cells to osteoblasts. Through RNA sequencing, RT-PCR and western blot, we found increased expression of FGFR3. Through co-immunoprecipitation assays and immunofluorescence staining, we revealed that hFGF23(A12D) activated the mitogen-activated protein kinase signalling pathway through interactions with the intracellular domain of FGFR3. In summary, we determined the mechanisms of hFGF23(A12D) involved in osteoblast generation and formation which is specifically due to its interaction with FGFR3.