Runx1促进BMP9诱导的间充质干细胞MEFs的成骨分化

doi:10.13523/j.cb.20170302

中国生物工程杂志

2017, Vol. 37

Issue (3): 10-17 DOI: 10.13523/j.cb.20170302

研究报告

Runx1促进BMP9诱导的间充质干细胞MEFs的成骨分化

吉彩霞, 刘晓骅, 徐丽, 董超群, 罗进勇

重庆医科大学检验医学院重庆医科大学检验医学教育部重点实验室重庆市重点实验室重庆 400016

Runx1 Enhances BMP9-induced Osteogenic Differentiation in Mesenchymal Stem Cell Line MEFs

JI Cai-xia, LIU Xiao-hua, XU Li, DONG Chao-qun, LUO Jin-yong

Key Laboratory of Diagnostic Medicine Designated by the Chinese Ministry of Education, Chongqing Medical University, Chongqing 400016, China

全文: PDF(1354 KB) HTML

摘要：

目的：研究Runx1在骨形态发生蛋白9（bone morphogenetic proteins 9，BMP9）诱导小鼠胚胎成纤维细胞（mouse embryonic fibroblasts，MEFs）成骨分化中的作用。方法：用BMP9腺病毒感染MEFs细胞，利用RT-PCR和Western blot分别在mRNA和蛋白质水平检测Runx1的内源性表达；构建过表达Runx1的重组腺病毒Ad-Runx1，并在mRNA和蛋白质水平验证Ad-Runx1的效果；用Ad-Runx1和BMP9条件培养基共处理MEFs，检测成骨早期指标碱性磷酸酶（ALP）染色和活性，茜素红S染色检测成骨晚期指标钙盐沉积；RT-PCR和Western blot分别检测成骨关键转录因子Runx2在mRNA和蛋白质水平的变化。结果：Ad-BMP9处理MEFs细胞后，可使Runx1在mRNA和蛋白质水平表达上调；构建的Ad-Runx1处理MEFs后，可使Runx1在mRNA和蛋白质水平表达上调；Ad-Runx1处理BMP9诱导的MEFs细胞后，增强了ALP活性和钙盐沉积以及Runx2在mRNA和蛋白质水平的表达。结论：Runx1可以促进BMP9诱导的间充质干细胞MEFs的成骨分化。

关键词： MEFs; 成骨分化; 骨形态发生蛋白9; Runx1

Abstract:

Objective:To investigate the role of Runx1 gene on BMP9-incuced osteogenic differentiation of Mouse embryonic fibroblasts (MEFs). Methods:MEFs were infected with Ad-BMP9 and then the expression of endogenous Runx1 at the mRNA and protein level was determined by RT-PCR and Western blot.The recombinant adenovirus Ad-Runx1 was constructed and its expression was validated at the mRNA and protein level. MEFs cells were treated with Ad-Runx1 or/and BMP9, the ALP activity was detected by quantitative and staining assay, calcium deposition was detected by Alizarin Red S staining. Runx2, as a core osteogenic transcription factor, was detected by RT-PCR and Western blot at the mRNA and protein level. Result:The expression of Runx1 was up-regulated by BMP9 at the mRNA and protein level in MEFs cells; Ad-Runx1 could enhance the expression of Runx1 at the mRNA and protein level. Runx1 can increase ALP activity and calcium deposition of MEFs cells induced by BMP9,and promote then expressions of Runx2 at the mRNA and protein level. Conclusion:BMP9-induced osteogenic differentiation is partially improved by Runx1 in mesenchymal stem cell line MEFs.

Key words: Runx1 Osteogenic differentiation MEFs BMP9

收稿日期: 2016-08-15 出版日期: 2017-03-25

ZTFLH:

Q813

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: 20182937@qq.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:

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.

JI Cai-xia, LIU Xiao-hua, XU Li, DONG Chao-qun, LUO Jin-yong. Runx1 Enhances BMP9-induced Osteogenic Differentiation in Mesenchymal Stem Cell Line MEFs. China Biotechnology, 2017, 37(3): 10-17.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170302 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I3/10

[1] Arthur A, Zannettino A, Gronthos S. The therapeutic applications of multipotential mesenchymal/stromal stem cells in skeletal tissue repair. J Cell Physiol,2009, 218(2):237-245.
[2] Molofsky A V, Pardal R, Morrison S J. Diverse mechanisms regulate stem cell self-renewal. Curr Opin Cell Biol,2004, 16(6):700-707.
[3] Myers T J, Granero-Molto F, Longobardi L, et al. Mesenchymal stem cells at the intersection of cell and gene therapy. Expert Opin Biol Ther, 2010, 10(12):1663-1679.
[4] Huang E, Bi Y, Jiang W, et al. Conditionally immortalized mouse embryonic fibroblasts retain proliferative activity without compromising multipotent differentiation potential. PLoS One, 2012, 7(2):e32428.
[5] Luu H H, Song W X, Luo X, et al. Distinct roles of bone morphogenetic proteins in osteogenic differentiation of mesenchymal stem cells. J Orthop Res, 2007, 25(5):665-677.
[6] Kang Q, Sun M H, Cheng H, et al. Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery. Gene Ther, 2004, 11(17):1312-1320.
[7] Lamplot J D, Qin J, Nan G, et al. BMP9 signaling in stem cell differentiation and osteogenesis. Am J Stem Cells, 2013, 2(1):1-21.
[8] Liu C, Weng Y, Yuan T, et al. CXCL12/CXCR4 signal axis plays an important role in mediating bone morphogenetic protein 9-induced osteogenic differentiation of mesenchymal stem cells. Int J Med Sci,2013, 10(9):1181-1192.
[9] Hojo H, Ohba S, He X, et al. Sp7/osterix is restricted to bone-forming vertebrates where It acts as a Dlx Co-factor in osteoblast specification. Dev Cell,2016, 37(3):238-253.
[10] Yoshida C A, Komori T. Role of Runx proteins in chondrogenesis. Crit Rev Eukaryot Gene Expr, 2005, 15(3):243-254.
[11] Chen C, Tang Z, Song Q, et al. Downregulated microRNA-23b promotes BMP9-mediated osteogenesis in C2C12 myoblast cells by targeting Runx2. Mol Med Rep,2016, 13(3):2492-2498.
[12] Wang J, Wang X, Holz J D, et al. Runx1 is critical for PTH-induced onset of mesenchymal progenitor cell chondrogenic differentiation. PLoS One,2013, 8(9):e74255.
[13] Wu M, Li C, Zhu G, et al. Deletion of core-binding factor beta (Cbfbeta) in mesenchymal progenitor cells provides new insights into Cbfbeta/Runxs complex function in cartilage and bone development. Bone,2014, 65:49-59.
[14] Loi F, Cordova L A, Pajarinen J, et al. Inflammation, fracture and bone repair. Bone,2016, 86:119-130.
[15] Saito T, Ohba S, Yano F, et al. Runx1 and Runx3 are downstream effectors of nanog in promoting osteogenic differentiation of the mouse mesenchymal cell line C3H10T1/2. Cell Reprogram, 2015, 17(3):227-234.
[16] Long L, Ormiston M L, Yang X, et al. Selective enhancement of endothelial BMPR-II with BMP9 reverses pulmonary arterial hypertension. Nat Med,2015, 21(7):777-785.
[17] Huang J, Yuan S X, Wang D X, et al. The role of COX-2 in mediating the effect of PTEN on BMP9 induced osteogenic differentiation in mouse embryonic fibroblasts. Biomaterials, 2014, 35(36):9649-9659.
[18] Smith N, Dong Y, Lian J B, et al. Overlapping expression of Runx1(Cbfa2) and Runx2(Cbfa1) transcription factors supports cooperative induction of skeletal development. J Cell Physiol,2005, 203(1):133-143.
[19] Wang Y, Belflower R M, Dong Y F, et al. Runx1/AML1/Cbfa2 mediates onset of mesenchymal cell differentiation toward chondrogenesis. J Bone Miner Res, 2005, 20(9):1624-1636.
[20] Yamashiro T, Wang X P, Li Z, et al. Possible roles of Runx1 and Sox9 in incipient intramembranous ossification. J Bone Miner Res, 2004, 19(10):1671-1677.
[21] Lu F, Gao J H. High efficient generation of recombinant adenovirus containing human Fas gene using a method of homologous recombination in bacteria. Chin Med J, 2004, 84(7):559-563.
[22] van Caam A, Madej W, Thijssen E, et al. Expression of TGFbeta-family signalling components in ageing cartilage:age-related loss of TGFbeta and BMP receptors. Osteoarthritis Cartilage, 2016, 24(7):1235-1245.
[23] Xu D J, Zhao Y Z, Wang J, et al. Smads, p38 and ERK1/2 are involved in BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells. BMB Rep,2012, 45(4):247-252.

[1]	朱颖,范梦恬,李具琼,陈彬,张盟浩,吴静红,施琼. 趋化因子受体CX3CR1调控人主动脉瓣膜间质细胞成骨分化的作用研究 ^*[J]. 中国生物工程杂志, 2019, 39(8): 7-16.
[2]	程瑜,施琼,安利钦,范梦恬,皇改改,翁亚光. **BMP7基因沉默抑制钙盐诱导猪主动脉瓣膜间质细胞成骨分化 ^***[J]. 中国生物工程杂志, 2019, 39(5): 63-71.
[3]	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.
[4]	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.
[5]	刘晓骅, 吉彩霞, 徐丽, 董超群, 罗进勇. Hmox1促进BMP9诱导C3H10T1/2细胞向成骨分化[J]. 中国生物工程杂志, 2017, 37(4): 33-39.
[6]	曹俊杰, 李爱芳, 卫亚琳, 廉静, 唐敏. Notch信号参与BMP4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, 2017, 37(4): 48-55.
[7]	徐丽, 吉彩霞, 刘晓骅, 喻婷婷, 罗进勇. DLX1对BMP9诱导的间充质干细胞C3H10T1/2成骨分化的影响[J]. 中国生物工程杂志, 2017, 37(10): 8-15.
[8]	李娅莎, 刘星, 毕杨, 杨珂, 赵丽, 龚梦嘉, 郭绮. BMP9诱导人脐带间充质干细胞体内外成骨分化的作用研究[J]. 中国生物工程杂志, 2016, 36(5): 20-26.
[9]	宋琪玲, 施琼, 陈楚, 唐祖川, 刘红霞, 周一青, 张汝益, 严树涓, 翁亚光. MiR-21协同BMP9促进间充质干细胞C3H10T1/2成骨分化[J]. 中国生物工程杂志, 2016, 36(2): 22-29.
[10]	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.
[11]	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.
[12]	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.
[13]	赵艳芳, 宋涛, 刘跃亮, 罗进勇. RUNX2对BMP9诱导的间充质干细胞C3H10T1/2成骨分化的影响[J]. 中国生物工程杂志, 2013, 33(2): 21-26.
[14]	李宝林, 白慧丽, 张汝益, 严树涓, 何方, 杨丹丹, 刘晨, 施琼. miR-30a抑制BMP9诱导间充质干细胞C3H10T1/2的成骨分化[J]. 中国生物工程杂志, 2013, 33(11): 14-20.
[15]	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.

Viewed

Full text

Abstract

Cited

Shared

Discussed