上海交通大学学报（医学版）2026，Vol.46Issue(7)：857-867,11.DOI:10.3969/j.issn.1674-8115.2026.07.004
细胞因子信号抑制因子3对小鼠颅顶成骨前体细胞成骨分化的影响
Effect of suppressor of cytokine signaling 3 on osteogenic differentiation of mouse calvarial osteoblast precursors
摘要
Abstract
Objective·To construct a stable suppressor of cytokine signaling 3(Socs3)-knockdown MC3T3-E1 mouse calvarial osteoblast precursor cell line and to investigate the role of SOCS3 in the osteogenic differentiation of calvarial osteoblast precursors.Methods·The expression levels of SOCS3 in MC3T3-E1 cells after osteogenic induction were detected by real-time quantitative PCR(RT-qPCR)and Western blotting.Two pairs of short hairpin RNAs(shRNAs)specifically targeting the Socs3 gene were designed,and lentiviral vectors for Socs3 knockdown were constructed.MC3T3-E1 cells were then infected with the packaged lentiviruses,followed by puromycin screening to obtain MC3T3-E1 cell lines with stable Socs3 knockdown.Knockdown efficiency was verified by RT-qPCR and Western blotting.The effect of SOCS3 on the proliferation capacity of MC3T3-E1 cells was assessed using a CCK-8 assay.After osteogenic differentiation for 7 d,alkaline phosphatase(ALP)staining and RT-qPCR detection of osteogenesis-related genes were performed.After 14 d of osteogenic differentiation,alizarin red S(ARS)staining was performed to assess the osteogenic differentiation capacity of the stable knockdown cell lines.Results·RT-qPCR and Western blotting showed that both Socs3 mRNA and SOCS3 protein levels were elevated at 4 d and 7 d after osteogenic induction compared with those before induction,and the levels at 7 d were higher than those at 4 d(both P<0.001).In MC3T3-E1 stable cell lines infected with sh Socs3-1 and sh Socs3-2 lentivirus,the expression levels of Socs3 mRNA and SOCS3 protein were significantly inhibited(P<0.001),indicating the successful establishment of stable Socs3-knockdown MC3T3-E1 cell lines.The CCK-8 assay showed that Socs3 knockdown inhibited the proliferation capacity of MC3T3-E1 cells at 1 d,4 d,and 7 d of culture(P<0.001).ALP staining demonstrated reduced ALP activity in the Socs3-knockdown MC3T3-E1 cell lines after 7 d of osteogenic induction(P<0.001).ARS staining indicated that Socs3 knockdown inhibited mineralized nodule formation in MC3T3-E1 cells(P<0.001).RT-qPCR detection of osteogenic marker genes indicated that the expression levels of runt-related transcription factor 2(Runx2),alkaline phosphatase(Alp),osterix(Osx),α1 type Ⅰ collagen(Col1a1),secreted phosphoprotein 1(Spp1),and bone γ-carboxyglutamate protein(Bglap)were significantly reduced in the Socs3-knockdown group(all P<0.001).Conclusion·SOCS3 expression increases during the osteogenic differentiation of MC3T3-E1 cells.Knockdown of Socs3 can inhibit both the proliferation and osteogenic differentiation capacities of MC3T3-E1 cells.关键词
细胞因子信号抑制因子3/MC3T3-E1细胞/颅顶成骨前体细胞/成骨分化Key words
suppressor of cytokine signaling 3(SOCS3)/MC3T3-E1 cell/calvarial osteoblast precursor/osteogenic differentiation分类
医药卫生引用本文复制引用
崔怡雯,孙思远,黄紫晗,代庆刚,江凌勇..细胞因子信号抑制因子3对小鼠颅顶成骨前体细胞成骨分化的影响[J].上海交通大学学报（医学版）,2026,46(7):857-867,11.基金项目
国家重点研发计划(2024YFC2510700) （2024YFC2510700）
国家自然科学基金(82430032,825B2029,825B1014) （82430032,825B2029,825B1014）
上海市科技创新行动计划国际科技合作项目/政府间国际科技合作项目(23410713600) （23410713600）
重庆市科卫联合医学科研项目开放项目(2026KFXM032) （2026KFXM032）
上海交通大学医学院"双百人"项目(20221809). National Key Research and Development Program of China(2024YFC2510700) （20221809）
National Natural Science Foundation of China(82430032,825B2029,825B1014) （82430032,825B2029,825B1014）
Shanghai Science and Technology Innovation Action Plan-International Science and Technology Cooperation Program(23410713600) （23410713600）
Chongqing Municipal Science and Health Joint Medical Research(2026KFXM032) （2026KFXM032）
"Two-hundred Talents"Program of Shanghai Jiao Tong University School of Medicine(20221809). （20221809）
