Molecular Mechanisms Study of WNT3A Promoting Osteoblasts Proliferation and Differentiation by Binding and Stabilizing FZD2 and Activating Wnt Signaling Pathway
CUI Yongjian, LI Yan, WANG Qiaomei, et al
The Sixth Affiliated Hospital of Xinjiang Medical University, Xinjiang Urumqi 830002, China
Abstract：Objective: To investigate the effects and the molecular mechanisms of ligand WNT3A on osteoblasts' bone formation by stabilizing and activating FZD2 (Frizzled2).Methods: Twenty-four female C57BL/6J mice aged 6~8 week were randomly divided into 4 groups: control group, sham group, ovariectomy-induced osteoporosis (OP) mouse model group (OVX group), and OVX+estradiol treatment group (OVX+E2 treatment group), with 6 mice in each group. The OVX mouse model was established. The expression levels of WNT3A, FZD2, Active-β-Catenin, β-Catenin, ALP and Runx2, and phosphorylated (p-) STAT3, STAT3, P-JAK2 and JAK2 in the tibia tissues of mouse hind limbs were determined by Western blot. The proliferation ability of mouse embryonic osteoblast MC3T3-E1 cells were determined by CCK-8 assay. Adenovirus-shRNA-FZD2 was used to knockdown FZD2 in MC3T3-E1 cells. Co-Immunoprecipitation (co-IP) was used for detecting the direct binding of FZD2 to ubiquitin (Ub) before and after MC3T3-E1 cells treated with WNT3A. Results: Compared with the OVX group, the expression levels of WNT3A, FZD2, Active-β-Catenin, β-Catenin, ALP, and Runx2 in the tibia tissue of mice in the OVX+E2 treatment group were up-regulated (P<0.05). Compared with Control group, in the WNT3A treatment group the expression levels of FZD2, Active-β-Catenin, β-Catenin, ALP, and Runx2 in MC3T3-E1 cells were increased (P<0.05); and the cell proliferation ability was increased (P<0.05). Compared with WNT3A treatment group, in the WNT3A treatment+Adv-shRNA-FZD2 group, the expression levels of FZD2, Active-β-Catenin, β-Catenin, ALP, and Runx2 were decreased (P<0.05); the phosphorylation levels of p-STAT3 and p-JAK2 were both decreased (P<0.05); the proliferation ability was decreased (P<0.05). There were no significant differences in the expression levels of STAT3 and JAK2 between the two groups (P>0.05). In the IP: Ub group, compared with WNT3A-treated (-) cells, the expression level of FZD2 was increased in WNT3A-treated (+) cells (P<0.05). Conclusion: The bone anabolic activity of WNT3A is mediated by binding and stabilizing FZD2 to activate the Wnt/β-Catenin signaling pathway.
崔永建, 李艳, 王巧梅, 唐庆. WNT3A结合并稳定FZD2激活Wnt通路促进成骨细胞增殖和分化的分子机制研究[J]. 河北医学, 2024, 30(3): 353-358.
CUI Yongjian, LI Yan, WANG Qiaomei, et al. Molecular Mechanisms Study of WNT3A Promoting Osteoblasts Proliferation and Differentiation by Binding and Stabilizing FZD2 and Activating Wnt Signaling Pathway. HeBei Med, 2024, 30(3): 353-358.
[1] Compston JE,McClung MR,Leslie WD.Osteoporosis[J].Lancet,2019,393(10169):364-376. [2] 焦楷磊,张宏光,夏维波,等.全国中老年骨质疏松症分级健康管理平台的设计与应用[J].中国骨质疏松杂志,2023,29(7):971-975. [3] Eastell R,Rosen CJ,Black DM,et al.Pharmacological management of osteoporosis in postmenopausal women:an endocrine society* clinical practice guideline[J].Clin Endocrinol Metab,2019,104(5):1595-1622. [4] Brown JP.Long-term treatment of postmenopausal osteoporosis[J].Endocrinol Metab (Seoul),2021,36(3):544-552. [5] Li J,Chen X,Lu L,et al.The relationship between bone marrow adipose tissue and bone metabolism in postmenopausal osteoporosis[J].Cytokine Growth Factor Rev,2020,(52):88-98. [6] Maeda K,Kobayashi Y,Koide M,et al.The regulation of bone metabolism and disorders by wnt signaling[J].Int Mol Sci,2019,20(22):5525. [7] Yu F,Chang J,Li J,et al.Protective effects of oridonin against osteoporosis by regulating immunity and activating the Wnt3a/β-catenin/VEGF pathway in ovariectomized mice[J].Int Immunopharmacol,2023(118):110011. [8] Zhu X,Xu M,Leu NA,et al.FZD2 regulates limb development by mediating β-catenin-dependent and -independent Wnt signaling pathways[J].Dis Model Mech,2023,16(3):49876. [9] Tosa I,Yamada D,Yasumatsu M,et al.Postnatal Runx2 deletion leads to low bone mass and adipocyte accumulation in mice bone tissues[J].Biochem Biophys Res Commun,2019,516(4):1229-1233. [10] Li X,Liu D,Li J,et al.Wnt3a involved in the mechanical loading on improvement of bone remodeling and angiogenesis in a postmenopausal osteoporosis mouse model[J].FASEB,2019,33(8):8913-8924. [11] Dijksterhuis JP,Baljinnyam B,Stanger K,et al.Systematic mapping of WNT-FZD protein interactions reveals functional selectivity by distinct WNT-FZD pairs[J].Biol Chem,2019,290(11):6789-6798.
2024, Vol. 30 
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