Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2015, Vol. 35 Issue (10): 20-26    DOI: 10.13523/j.cb.20151003
研究报告     
SATB2促进BMP9诱导C2C12细胞的成骨分化
冯巧灵, 王玉凤, 刘晓骅, 吉彩霞, 罗进勇
重庆医科大学检验医学院 临床检验诊断学教育部重点实验室 重庆市重点实验室 重庆 400016
SATB2 Promotes Bone Morphogenetic Protein 9-induced Osteogenic Differentiation of Mesenchymal Stem Cells
FENG Qiao-ling, WANG Yu-feng, LIU Xiao-hua, JI Cai-xia, LUO Jin-yong
Key laboratory of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China
 全文: PDF(1359 KB)   HTML
摘要:

目的:研究特异AT序列结合蛋白2(Special AT-rich sequence-binding protein 2,SATB2)在骨形态发生蛋白9(bone morphogenetic proteins 9,BMP9) 诱导的间充质干细胞(mesenchymal stem cells,MSCs)C2C12成骨分化中的作用。方法:首先用Ad-BMP9感染C2C12细胞,RT-PCR检测 SATB2 在基因水平上的表达变化,Western blot检测SATB2在蛋白水平上的变化;构建过表达SATB2重组腺病毒Ad-SATB2,感染C2C12细胞后,Western blot验证Ad-SATB2表达情况;用Ad-SATB2处理BMP9诱导的C2C12细胞,碱性磷酸酶(ALP)活性和染色检测成骨早期ALP的变化,茜素红S染色检测成骨晚期指标钙盐沉积的变化。结果:Ad-BMP9 处理C2C12细胞后,比对照组SATB2在基因水平和蛋白水平上表达量上调;Ad-SATB2可以在细胞中成功表达SATB2蛋白;用Ad-SATB2处理BMP9诱导的C2C12细胞后,ALP以及钙盐的沉积与对照比较均上调。结论:SATB2可以促进BMP9诱导的间充质干细胞C2C12的成骨分化。

关键词: 特异AT序列结合蛋白2骨形态发生蛋白9C2C12细胞    
Abstract:

Objective: To analyze the effect of SATB2 on BMP9-induced osteogenic differentiation of C2C12 mesenchymal stem cells. Methods: C2C12 cells were treated with ad-BMP9, and then the expressions of SATB2 at the gene and protein level were detected with RT-PCR and Western blot, respectively. Afterwards C2C12 cells were treated with ad-SATB2 or/and BMP9, then ALP activity was detected by quantitative and staining assay, and calcium deposition was detected by Alizarin Red S staining. Results: BMP9 can enhance the expression of SATB2 at the gene level and at the protein level in C2C12 cells. Both ALP activity and calcium deposition of C2C12 cells treated with adenovirus SATB2 or/and BMP9 expressed higher than those of controls. Conclusions: SATB2 can promote BMP9-induced osteogenic differentiation of C2C12 mesenchymal stem cells

Key words: Bone morphogenetic protein 9    C2C12 cells    Special AT-rich sequence binding protein-2
收稿日期: 2015-05-24 出版日期: 2015-10-25
ZTFLH:  Q254  
基金资助:

国家自然科学基金资助项目(31071304,81272006)

通讯作者: 罗进勇     E-mail: qiaoling1026@foxmail.com
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  

引用本文:

冯巧灵, 王玉凤, 刘晓骅, 吉彩霞, 罗进勇. SATB2促进BMP9诱导C2C12细胞的成骨分化[J]. 中国生物工程杂志, 2015, 35(10): 20-26.

FENG Qiao-ling, WANG Yu-feng, LIU Xiao-hua, JI Cai-xia, LUO Jin-yong. SATB2 Promotes Bone Morphogenetic Protein 9-induced Osteogenic Differentiation of Mesenchymal Stem Cells. China Biotechnology, 2015, 35(10): 20-26.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20151003        https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I10/20

[1] Pittenger M F, Mackay A M, Beck S C, et al. Multilineage potential of adult human mesenchymal stem cells. Science, 1999, 284(5411):143-147.
[2] Horwitz E M, Prockop D J, Fitzpatrick L A, et al. Transplantability and therapeutic effects of bone marrow-derived mesenchymal cells in children with osteogenesis imperfecta. Nat Med, 1999, 5(3):309-313.
[3] Jiang Y, Jahagirdar B N, Reinhardt R L, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature, 2002, 418(6893):41-49.
[4] Tseng Y H, Kokkotou E, Schulz T J, et al. New role of bone morphogenetic protein 7 in brown adipogenesis and energy expenditure. Nature, 2008, 454(7207):1000-1004.
[5] Cheng H, Jiang W, Phillips F M, et al. Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs). J Bone Joint Surg Am, 2003, 85-A(8):1544-1552.
[6] Govender S, Csimma C, Genant H K, et al. Recombinant human bone morphogenetic protein-2 for treatment of open tibial fractures: a prospective, controlled, randomized study of four hundred and fifty patients. J Bone Joint Surg Am, 2002, 84-A(12):2123-2134.
[7] Luo J, Tang M, Huang J, et al. TGFbeta/BMP type I receptors ALK1 and ALK2 are essential for BMP9-induced osteogenic signaling in mesenchymal stem cells. J Biol Chem, 2010, 285(38):29588-29598.
[8] Sinha K M, Zhou X. Genetic and molecular control of osterix in skeletal formation. J Cell Biochem, 2013, 114(5):975-984.
[9] Lee K S, Kim H J, Li Q L, et al. Runx2 is a common target of transforming growth factor beta1 and bone morphogenetic protein 2, and cooperation between Runx2 and Smad5 induces osteoblast-specific gene expression in the pluripotent mesenchymal precursor cell line C2C12. Mol Cell Biol, 2000, 20(23):8783-8792.
[10] Dobreva G, Chahrour M, Dautzenberg M, et al. SATB2 is a multifunctional determinant of craniofacial patterning and osteoblast differentiation. Cell, 2006, 125(5):971-986.
[11] Zhang P, Men J, Fu Y, et . Contribution of SATB2 to the stronger osteogenic potential of bone marrow stromal cells from craniofacial bones. Cell Tissue Res, 2012, 350(3):425-437.
[12] Savarese F, Davila A, Nechanitzky R, et al. Satb1 and Satb2 regulate embryonic stem cell differentiation and Nanog expression. Genes Dev, 2009, 23(22):2625-2638.
[13] Asanoma K, Kubota K, Chakraborty D, et al. SATB homeobox proteins regulate trophoblast stem cell renewal and differentiation. J Biol Chem, 2012, 287(3):2257-2268.
[14] Tu Q, Valverde P, Li S, et al. Osterix overexpression in mesenchymal stem cells stimulates healing of critical-sized defects in murine calvarial bone. Tissue Eng, 2007, 13(10):2431-2440.
[15] He T C, Zhou S, Da C L, et al. A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci U S A, 1998, 95(5): 2509-2514.
[16] Mendez-Ferrer S, Michurina T V, Ferraro F, et al. Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature, 2010, 466(7308):829-834.
[17] Chambers I, Colby D, Robertson M, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell, 2003, 113(5):643-655.
[18] Chang H Y. Anatomic demarcation of cells: Genes to patterns. Science, 2009, 326(5957):1206-1207.
[19] Dong W, Zhang P, Fu Y, et al. Roles of SATB2 in site-specific stemness, autophagy and senescence of bone marrow mesenchymal stem cells. J Cell Physiol, 2015, 230(3):680-690.
[20] Gong Y, Qian Y, Yang F, et al. Lentiviral-mediated expression of SATB2 promotes osteogenic differentiation of bone marrow stromal cells in vitro and in vivo. Eur J Oral Sci, 2014, 122(3):190-197.
[21] Asanoma K, Kubota K, Chakraborty D, et al. SATB homeobox proteins regulate trophoblast stem cell renewal and differentiation. J Biol Chem, 2012, 287(3):2257-2268.
[22] Chang H Y, Chi J T, Dudoit S, et al. Diversity, topographic differentiation, and positional memory in human fibroblasts. Proc Natl Acad Sci USA, 2002, 99(5):12877-12882.

[1] 郑妍,姚欢,杨珂. SFRP5抑制BMP9诱导人脐带间充质干细胞成骨分化的实验研究 *[J]. 中国生物工程杂志, 2018, 38(7): 7-13.
[2] 刘红霞, 施琼, 周一青, 安利钦, 严树涓, 张汝益, 翁亚光. 过表达miR-155抑制BMP9诱导间充质干细胞C3H10T1/2成骨分化[J]. 中国生物工程杂志, 2017, 37(5): 9-18.
[3] 吉彩霞, 刘晓骅, 徐丽, 董超群, 罗进勇. Runx1促进BMP9诱导的间充质干细胞MEFs的成骨分化[J]. 中国生物工程杂志, 2017, 37(3): 10-17.
[4] 唐大刚, 王淼, 张艳亮, 王孝林, 李贵强, 罗小辑. Hey1 基因参与调控BMP9诱导的C3H10T1/2细胞成骨分化[J]. 中国生物工程杂志, 2015, 35(6): 14-20.
[5] 李丽, 蒙秋蓉, 郭琦, 王岚, 商蕾, 欧欣颖, 罗进勇. Shh信号参与调控BMP9诱导的间充质干细胞成骨分化[J]. 中国生物工程杂志, 2014, 34(9): 9-15.
[6] 谢佳瑛, 胥文春, 徐道晶, 张晓艳, 唐敏. Notch信号参与BMP9诱导的间充质干细胞成骨分化及其机制的初步探讨[J]. 中国生物工程杂志, 2012, 32(11): 14-22.
[7] 谢佳瑛, 胥文春, 徐道晶, 张晓艳, 唐敏. Notch信号参与BMP9诱导的间充质干细胞成骨分化及其机制的初步探讨[J]. 中国生物工程杂志, 2012, 32(11): 14-22.
[8] 徐道晶, 王锦, 何娟文, 胡晶, 翁亚光, 罗进勇. p38蛋白激酶参与BMP9诱导的C3H10T1/2细胞成骨分化[J]. 中国生物工程杂志, 2011, 31(5): 15-21.
[9] 徐道晶 王锦 何娟文 胡晶 翁亚光 罗进勇. p38蛋白激酶参与BMP9诱导的C3H10T1/2细胞成骨分化[J]. 中国生物工程杂志, 2011, 31(05): 0-0.