›› 2011, Vol. 31 ›› Issue (12): 1661-.doi: 10.3969/j.issn.1674-8115.2011.12.001

• 论著(基础研究) •    下一篇

MSCs成骨分化中BMP2对成骨转录因子SATB2表达的调控作用

左炽健1, 剌 婷1, 张 宁1, 戴尅戎1,2, 张晓玲1,2   

  1. 1.中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所骨科细胞与分子生物学实验室, 上海 200025; 2.上海市骨科内植物重点实验室 上海交通大学医学院附属第九人民医院骨科, 上海 200011
  • 出版日期:2011-12-28 发布日期:2012-01-04
  • 通讯作者: 张晓玲, 电子信箱: xlzhang@sibs.ac.cn。
  • 作者简介:左炽健(1985—), 男, 硕士生;电子信箱: jian2005a@sina.com。
  • 基金资助:

    国家自然科学基金(30871435);上海市科委国际合作基金(09540703600);上海市教委曙光计划(10SG22);上海教委重点学科建设基金(J50206)

Regulation of expression of osteoblast transcription factor SATB2 by BMP2 in osteoblastic differentiation of MSCs

ZUO Chi-jian1, LA Ting1, ZHANG Ning1, DAI Ke-rong1,2, ZHANG Xiao-ling1,2   

  1. 1.Laboratory of Orthopedic Cellular and Molecular Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences &|Shanghai Jiaotong University School of Medicine, Shanghai 200025, China;2.Shanghai Key Laboratory of Orthopaedic Implant, Department of Orthopaedics, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2011-12-28 Published:2012-01-04
  • Supported by:

    National Natural Science Foundation of China, 30871435;Shanghai Science and Technology Committee Foundation, 09540703600;Shanghai Education Committee Foundation, 10SG22, J50206

摘要:

目的 探索间充质干细胞(MSCs)成骨分化中骨形态发生蛋白2 (BMP2)对成骨转录因子SATB2 表达的调控作用。方法 体外培养小鼠间充质细胞系C2C12,腺病毒介导的BMP2 (Adv-BMP2) 诱导其向成骨细胞分化,建立并验证C2C12细胞成骨分化细胞模型。Real-Time PCR和Western blotting 分别检测C2C12细胞成骨分化过程中经不同浓度Adv-BMP2处理不同时间时SATB2 mRNA和SATB2蛋白表达;以经相应浓度Adv-β-Gal处理细胞作对照。结果 经150 pfu/cell Adv-BMP2处理C2C12细胞5 d后,成骨细胞标志基因Ⅰ型胶原、骨唾液酸蛋白和骨钙素表达以及碱性磷酸酶活性均显著增加,MSCs成骨分化模型构建成功。150 pfu/cell Adv-BMP2诱导C2C12细胞成骨分化过程中,SATB2 mRNA和SATB蛋白表达随分化进程而增加;Adv-BMP2浓度为0~225 pfu/cell时,SATB2表达随Adv-BMP2浓度升高而增加。结论 BMP2 可调控SATB2的表达,从而影响MSCs成骨分化。

关键词: 骨形态发生蛋白2, 成骨转录因子, SATB2, 基因表达, 调控

Abstract:

Objective To investigate the role of bone morphogenetic protein 2 (BMP2) in the regulation of expression of osteoblast transcription factor SATB2 in osteoblastic differentiation of mesenchymal stem cells (MSCs). Methods Mesenchymal cell line C2C12 was stimulated by adenovirus-derived BMP2 (Adv-BMP2) for osteoblastic differentiation, and model of osteoblastic differentiation of C2C12 cells was established and verified. Real-Time PCR and Western blotting were employed to detect the expression of SATB2 mRNA and SATB2 protein respectively in osteoblastic differentiation treated by Adv-BMP2 for different time and by Adv-BMP2 of different concentrations. C2Cl2 cells treated with Adv-β-Gal were served as controls. Results After treatment by 150 pfu/cell Adv-BMP2 for 5 d, the expression of collagen type Ⅰ, bone sialoprotein and osteocalcin and the activity of alkaline phosphatase in C2C12 cells significantly increased, and the model of osteoblastic differentiation of C2Cl2 cells was established. The expression of SATB2 mRNA and SATB2 protein gradually increased with the progression of osteoblastic differentiation of C2Cl2 cells treated by 150 pfu/cell AdvBMP2, and the expression of SATB2 mRNA and SATB2 protein increased with the concentration increase of 0-225 pfu/cell Adv-BMP2. Conclusion BMP2 may influence osteoblastic differentiation of MSCs by regulating the expression of SATB2.

Key words: bone morphogenetic protein 2, osteoblast transcription factor, SATB2, gene expression, regulation