微阵列芯片分析成牙骨质细胞矿化过程中microRNA表达谱的差异

doi:10.13701/j.cnki.kqyxyj.2017.02.005

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (2): 135-139.DOI: 10.13701/j.cnki.kqyxyj.2017.02.005

微阵列芯片分析成牙骨质细胞矿化过程中microRNA表达谱的差异

王川¹, 廖海清¹, 曹正国^1,2*

1. 武汉大学口腔医学院口腔基础医学省部共建国家重点实验室培训基地和口腔生物医学教育部重点实验室湖北武汉 430079;
2. 武汉大学口腔医院牙周科湖北武汉 430079

收稿日期:2016-11-01 出版日期:2017-02-20 发布日期:2017-02-22
通讯作者: 曹正国,电话:027-87686212
作者简介:王川(1990~ ),男,河南人,硕士在读,研究方向为牙周病学。
基金资助:
国家自然科学基金(编号:81170933,81570946)湖北省自然科学基金面上项目(2015CFB259)

Differentially Expressed MicroRNA during Cementoblasts Mineralization Based Microarray.

WANG Chuan¹, LIAO Hai-qing¹, CAO Zheng-guo^1,2*.

1. The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST KLOS &
Key Laboratory for Oral Biomedical Engineering of Ministry of Education(KLOBME, School &
Hospital of Stomatology, Wuhan University, Wuhan 430079, China;
2. Department of Periodontology, School &
Hospital of Stomatology, Wuhan University, Wuhan 430079, China.

Received:2016-11-01 Online:2017-02-20 Published:2017-02-22

摘要/Abstract

摘要： 目的:诱导小鼠成牙骨质细胞矿化,研究在成牙骨质细胞矿化过程中microRNA表达谱的变化。方法:体外培养小鼠成牙骨质细胞系OCCM30,使用抗坏血酸、β-甘油磷酸钠和地塞米松诱导成牙骨质细胞矿化。实时荧光定量PCR(qPCR)检测碱性磷酸酶和骨钙蛋白表达量变化来验证矿化情况。应用miRNA microarray 芯片技术,分析成牙骨质细胞矿化过程中miRNA表达谱的变化。QPCR对芯片结果进行验证。结果:成功诱导了成牙骨质细胞的矿化。MiRNA microarray 芯片检测发现成牙骨质细胞矿化过程中miRNA表达谱出现明显改变,其中上调的有4个,下调的有39个。QPCR证明芯片结果准确可靠。结论:建立了小鼠成牙骨质细胞矿化的细胞模型,应用miRNA芯片发现小鼠成牙骨质细胞矿化过程中差异表达的miRNA,提示这些miRNA可能参与成牙骨质细胞矿化过程的调控。

关键词: 成牙骨质细胞, 矿化, microRNA, 微阵列芯片

Abstract: Objective: To establish the mineralization cell model of cementoblasts and to study the microRNA expression profile during the differentiation and mineralization of cementoblasts. Methods: Mice cementoblast cell line OCCM30 was cultured in vitro and the cementoblasts were differentiated and mineralized in the presence of ascorbic acid, β-glycerophosphate, and dexamethasone. Then, the mineralization related markers ALP and OC were assessed by qPCR. Finally, the differentiated miRNA expression profile during the differentiation and mineralization of cementoblasts was screened by miRNA microarray and the accuracy of the results was verified by qPCR. Results: Cementoblasts were successfully induced to differentiation and mineralization. MiRNA microarray showed that there were 4 miRNAs were up-regulated and 39 miRNAs were down-regulated during this process. QPCR confirmed the accuracy of miRNA microarray. Conclusion: The cementoblast mineralization model was established successfully. MiRNA microarray discovered that the miRNAs were differently expressed during the mineralization of cementoblasts. This indicated that these miRNAs may have certain roles during the differentiation and mineralization of cementoblasts.

Key words: Cementoblast , Mineralization , MicroRNA, Microarray

中图分类号:

R781.4

王川, 廖海清, 曹正国. 微阵列芯片分析成牙骨质细胞矿化过程中microRNA表达谱的差异[J]. 口腔医学研究, 2017, 33(2): 135-139.

WANG Chuan, LIAO Hai-qing, CAO Zheng-guo.. Differentially Expressed MicroRNA during Cementoblasts Mineralization Based Microarray.[J]. Journal of Oral Science Research, 2017, 33(2): 135-139.

参考文献

[1] Bosshardt, D.D.,K.A. Selvig,Dental cementum: the dynamic tissue covering of the root [J]. Periodontol 2000,1997,13(1)∶41-75
[2] Zeichner-David, M.,Regeneration of periodontal tissues: cementogenesis revisited [J]. Periodontol 2000,2006,41(2)∶196-217
[3] Grzesik, W.J.,A.S. Narayanan,Cementum and periodontal wound healing and regeneration [J]. Crit Rev Oral Biol Med,2002,13(6)∶474-84
[4] D'Errico, J.A.,R.L. MacNeil,T. Takata ,et al. Expression of bone associated markers by tooth root lining cells, in situ and in vitro [J]. Bone,1997,20(2)∶117-26
[5] Bosshardt, D.D. Are cementoblasts a subpopulation of osteoblasts or a unique phenotype [J]. J Dent Res,2005,84(5)∶390-406
[6] Bartel, D.P.,MicroRNAs: genomics, biogenesis, mechanism, and function [J]. Cell,2004,116(2)∶281-97
[7] Huang, C.,J. Geng,S. Jiang,MicroRNAs in regulation of osteogenic differentiation of mesenchymal stem cells [J]. Cell Tissue Res,2016:[Epub ahead of print]
[8] Yu, S.,Q. Geng,Q. Pan, et al. MiR-690, a Runx2-targeted miRNA, regulates osteogenic differentiation of C2C12 myogenic progenitor cells by targeting NF-kappaB p65 [J]. Cell Biosci,2016,6(1)∶10-16
[9] Huang, C.,J. Geng,X. Wei, et al. MiR-144-3p regulates osteogenic differentiation and proliferation of murine mesenchymal stem cells by specifically targeting Smad4 [J]. FEBS Lett,2016,590(6)∶795-807
[10] Li, J.,X. He,W. Wei, et al. MicroRNA-194 promotes osteoblast differentiation via downregulating STAT1 [J]. Biochem Biophys Res Commun,2015,460(2)∶482-8
[11] 胡芝爱,潘洁,李煜昱,等.DAPT对成牙骨质细胞增殖活性的影响[J].口腔医学研究,2016,32(2)∶117-121
[12] 邱雨,任嫒姝,庞琴,等.信号素3A对成牙骨质细胞系OCCM-30增殖和矿化功能影响的实验研究[J].口腔医学研究,2016,32(5)∶445-448
[13] Cao, Z.,H. Zhang,X. Zhou, et al. Genetic evidence for the vital function of Osterix in cementogenesis [J]. J Bone Miner Res,2012,27(5)∶1080-92
[14] Cao, Z.,R. Liu,H. Zhang, et al. Osterix controls cementoblast differentiation through downregulation of Wnt-signaling via enhancing DKK1 expression [J]. Int J Biol Sci,2015,11(3)∶335-44
[15] 蔡洁明,舒传继,杨伟民,等.miRNA-31和STAT-3在舌鳞状细胞癌中的表达及临床意义[J].口腔医学研究,2015,31(8)∶820-823
[16] 蒋少云,薛栋,邓嘉胤.小分子RNA在干细胞成骨及成软骨分化中的作用[J].口腔医学研究,2015,31(8)∶845-847
[17] 王鑫,王姗,孟琰,等.NSCL-P患者中异常表达microRNA与WNT家族的关联性研究[J].口腔医学研究,2016,32(5)∶470-473

微阵列芯片分析成牙骨质细胞矿化过程中microRNA表达谱的差异

Differentially Expressed MicroRNA during Cementoblasts Mineralization Based Microarray.

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	田子璐, 朱松. 诱导牙本质再矿化以堵塞牙本质小管的研究进展[J]. 口腔医学研究, 2021, 37(1): 11-14.
[2]	王岩松, 刘建林, 陈晓霞 ,白姝婷. Exendin4-PEGDA/HA矿化水凝胶的制备及体外成骨作用的研究[J]. 口腔医学研究, 2020, 36(8): 731-735.
[3]	岳阳丽, 马贵霞, 刘翠丽, 贺贵天. 生物活性玻璃促进牙本质再矿化的时间相关性体外研究[J]. 口腔医学研究, 2020, 36(12): 1137-1141.
[4]	王金凤, 车静怡, 刘会梅, 朱兴国, 孙文章, 潘爽. 黄瓜籽醇提取物对早期釉质龋再矿化的影响[J]. 口腔医学研究, 2020, 36(1): 65-69.
[5]	胡蝶,张凌琳. 口腔来源蛋白及多肽诱导牙釉质仿生矿化的研究进展[J]. 口腔医学研究, 2019, 35(6): 517-520.
[6]	宋金花, 刘宗仁, 刘文超, 张旭, 李彦秋. 羧甲基壳聚糖-溶菌酶纳米凝胶稳定无定形磷酸钙促进脱矿牙釉质再矿化[J]. 口腔医学研究, 2018, 34(8): 828-831.
[7]	王欣,张程,李天成,陶贵渝,段沛沛,邹淑娟. 一种新型牙骨质细胞系的矿化特性研究[J]. 口腔医学研究, 2018, 34(12): 1324-1328.
[8]	杨薇, 王健平, 刘鑫喆, 黄歆博, 李雪. 氟纳米羟基磷灰石对早期根面龋再矿化效果的实验研究[J]. 口腔医学研究, 2017, 33(2): 191-193.
[9]	梁向阳,孔晶晶,李春年,李安. 不同材料在碳酸饮料所致脱矿釉质再矿化中的作用[J]. 口腔医学研究, 2017, 33(12): 1274-1277.
[10]	王贵玲, 杨谛, 郭佳杰, 于雅琼, 仇丽鸿. MicroRNA 34a-5p对成骨细胞炎症反应及分化的影响[J]. 口腔医学研究, 2017, 33(11): 1139-1142.
[11]	邱雨 ,任嫒姝 ,庞琴 ,马小娟 ,付钢. 信号素3A对成牙骨质细胞系OCCM-30增殖和矿化功能影响的实验研究[J]. 口腔医学研究, 2016, 32(5): 445-448.
[12]	余淼,吴燕茹,韩雨亭,滕方君,黄亦青,王家伟. Ddit3在小鼠下颌第一磨牙牙胚不同时期的表达研究[J]. 口腔医学研究, 2016, 32(4): 331-334.
[13]	于莉,李淑慧,袁萍,赵璐,周春梅,吴佩玲. TNF-α对根尖乳头干细胞体外增殖及成牙成骨向分化能力的影响[J]. 口腔医学研究, 2016, 32(4): 343-346.
[14]	刘珊,张洁,朱玲新,彭彬. iRoot BP Plus对小鼠炎性骨破坏作用的研究[J]. 口腔医学研究, 2016, 32(2): 113-116.
[15]	胡芝爱,潘洁,李煜昱,周泽渊,黄鹂,邹淑娟. DAPT对成牙骨质细胞增殖活性的影响[J]. 口腔医学研究, 2016, 32(2): 117-121.