人牙髓干细胞成骨分化前后相关基因的表达分析

doi:10.13701/j.cnki.kqyxyj.2018.04.010

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (4): 375-379.DOI: 10.13701/j.cnki.kqyxyj.2018.04.010

人牙髓干细胞成骨分化前后相关基因的表达分析

林文珍, 高丽, 牛晨光, 胡戌琛, 苑克勇, 马瑞, 黄正蔚^*

上海交通大学医学院附属第九人民医院牙体牙髓病科;上海市口腔医学重点实验室/上海市口腔医学研究所,国家口腔疾病临床研究中心上海 200011

收稿日期:2017-09-23 出版日期:2018-04-28 发布日期:2018-04-25
通讯作者: 黄正蔚,E-mail:huangzhengwei@shsmu.edu.cn
作者简介:林文珍(1991~ ),女,浙江温州人,硕士在读,主要从事牙髓干细胞相关的研究。
基金资助:
国家自然科学基金(编号:81570964)上海市教育委员会高峰高原学科建设计划

Gene Expression of Human Dental Pulp Stem Cells during Osteogenic Differentiation.

LIN Wen-zhen, GAO Li, NIU Chen-guang, HU Xu-chen, YUAN Ke-yong, MA Rui, HUANG Zheng-wei^*

Department of Endodontics, Ninth People's Hospital, Shanghai JiaoTong University School of Medicine; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, Shanghai 200011, China.

Received:2017-09-23 Online:2018-04-28 Published:2018-04-25

摘要/Abstract

摘要： 目的:分析人牙髓干细胞(hDPSCs)成骨诱导后相关基因的表达变化。方法:使用改良组织块法体外培养hDPSCs并检测其多向分化能力。hDPSCs成骨诱导后对其进行基因表达谱芯片技术和实时荧光定量PCR分析相关基因的表达情况。结果:hDPSCs能够进行成骨细胞和脂肪细胞分化。基因表达谱芯片技术分析得出hDPSCs在成骨诱导前后有1284个基因上调或下调2倍以上。其中,骨调节蛋白(osteomodulin,OMD)基因转录上调约50倍。实时荧光定量PCR显示OMD基因在成骨诱导过程中转录呈逐渐增加趋势。结论:hDPSCs成骨分化涉及多个基因调控。OMD基因也参与其中,其表达变化具有时间规律性,与成骨过程呈正相关。

关键词: 人牙髓干细胞, 成骨分化, 骨调节蛋白

Abstract: Objective: To evaluate the differences in gene expression of human dental pulp stem cells (hDPSCs) after osteogenetic induction.. Methods: hDPSCs were cultured by explant method and the capability of multi-differentiation in hDPSCs was identified. The gene expressions were evaluated by microarray assay and quantitative real-time PCR after osteogenetic induction. Results: hDPSCs could differentiate into osteoblasts and adipocytes. The microarray assay revealed that the expression of 1284 genes were upregulated or downregulated more than twice after osteogenetic induction. Of these, the transcription of osteomodulin (OMD) gene was upregulated by fifty times. The results of qPCR indicated that the expression of OMD was gradually increased during osteogenetic induction. Conclusion: The osteogenetic differentiation of hDPSCs is regulated by multiple genes. OMD is one of these genes, and its expression is time-dependent and coincides with osteogenesis.

Key words: Human dental pulp stem cells, Osteogenetic differentiation, Osteomodulin

林文珍, 高丽, 牛晨光, 胡戌琛, 苑克勇, 马瑞, 黄正蔚. 人牙髓干细胞成骨分化前后相关基因的表达分析[J]. 口腔医学研究, 2018, 34(4): 375-379.

LIN Wen-zhen, GAO Li, NIU Chen-guang, HU Xu-chen, YUAN Ke-yong, MA Rui, HUANG Zheng-wei. Gene Expression of Human Dental Pulp Stem Cells during Osteogenic Differentiation.[J]. Journal of Oral Science Research, 2018, 34(4): 375-379.

参考文献

[1] Diomede F, Rajan TS, Gatta V, et al. Stemness maintenance properties in human oral stem cells after long-term passage [J]. Stem Cells Int, 2017, 2017∶5651287
[2] Gronthos S, Mankani M, Brahim J, et al. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo [J]. Proc Natl Acad Sci U S A, 2000, 97(25)∶13625-13630
[3] Xia XL, Brophy S, Zhou SM. Learning differentially expressed gene pairs in microarray data [J]. Stud Health Technol Inform, 2017, 235∶191-195
[4] Nakamura S, Yamada Y, Katagiri W, et al. Stem cell proliferation pathways comparison between human exfoliated deciduous teeth and dental pulp stem cells by gene expression profile from promising dental pulp [J]. J Endod, 2009, 35(11)∶1536-1542
[5] 高丽,江文欣,牛晨光,等.干细胞表面标志物在牙髓干细胞中的表达[J].上海口腔医学,2015,24(3)∶257-262
[6] Struys T, Moreels M, Martens W, et al. Ultrastructural and immunocytochemical analysis of multilineage differentiated human dental pulp- and umbilical cord-derived mesenchymal stem cells [J]. Cells Tissues Organs, 2011, 193(6)∶366-378
[7] 刘金凤,牛雪微,超博,等.牙髓再生的研究现状[J].口腔医学研究, 2017, 33(1)∶108-111
[8] Augello A, De Bari C. The regulation of differentiation in mesenchymal stem cells [J]. Hum Gene Ther, 2010, 21(10)∶1226-2238
[9] Liu J, Jin T, Chang S, et al. Matrix and TGF-beta-related gene expression during human dental pulp stem cell (DPSC) mineralization [J]. In Vitro Cell Dev Biol Anim, 2007, 43(3/4)∶120-128
[10] Datta P, Ghosh P, Ghosh K, et al .in vitro ALP and osteocalcin gene expression analysis and in vivo biocompatibility of N-methylene phosphonic chitosan nanofibers for bone regeneration [J]. J Biomed Nanotechnol, 2013, 9(5)∶870-879
[11] Sommarin Y, Wendel M, Shen Z, et al. Osteoadherin, a cell-binding keratan sulfate proteoglycan in bone, belongs to the family of leucine-rich repeat proteins of the extracellular matrix [J]. J Biol Chem, 1998, 273 (27)∶16723- 16729
[12] Ninomiya K, Miyamoto T, Imai J, et al. Osteoclastic activity induces osteomodulin expression in osteoblasts [J]. Biochem Biophys Res Commun, 2007, 362(2)∶460-466
[13] Tashima T, Nagatoishi S, Sagara H, et al. Osteomodulin regulates diameter and alters shape of collagen fibrils [J]. Biochem Biophys Res Commun, 2015, 463(3)∶292-296
[14] Rehn AP, Chalk AM, Wendel M. Differential regulation of osteoadherin (OSAD) by TGF-β1 and BMP-2 [J]. Biochem Biophys Res Commun, 2006, 349(3)∶1057-1064

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人牙髓干细胞成骨分化前后相关基因的表达分析

Gene Expression of Human Dental Pulp Stem Cells during Osteogenic Differentiation.

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