BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化

doi:10.19439/j.sjos.2017.05.003

上海口腔医学 ›› 2017, Vol. 26 ›› Issue (5): 476-483.doi: 10.19439/j.sjos.2017.05.003

BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化

包丽荣¹, 赵文青¹, 林田¹, 陆彦玲¹, 吴煜²

1.广西医科大学,广西南宁 530021；
2.广西医科大学附属口腔医院儿童口腔科,广西南宁 530021

收稿日期:2017-01-22 修回日期:2017-03-23 出版日期:2017-10-25 发布日期:2017-11-23
通讯作者: 吴煜,E-mail:wuyu0912@163.com
作者简介:包丽荣（1992-）,女,在读硕士研究生,E-mail:blr1992@163.com
基金资助:
广西科学基金项目（2011GXNSFA018216）; 广西医疗卫生适宜技术研究与开发项目（S201305-02）; 广西高校科学技术研究项目（2013YB042）

miRNA profile of the human dental pulp cells during odontoblast differentiation induced by BMP-2

BAO Li-rong¹, ZHAO Wen-qing¹, LIN Tian¹, LU Yan-ling¹, WU Yu²

1.Guangxi Medical University. Nanning 530021;
2. Department of Paediatric Dentistry, Affiliated Stomatological Hospital of Guangxi Medical University. Nanning 530021, Guangxi Zhuang Autonomous Region, China

Received:2017-01-22 Revised:2017-03-23 Online:2017-10-25 Published:2017-11-23

摘要/Abstract

摘要： 目的筛选出骨形态发生蛋白2（bone morphogenetic protein-2,BMP-2）诱导人牙髓细胞（human dental pulp cells,hDPCs）成牙本质向分化时差异表达的微小RNA(miRNAs)。方法以BMP-2诱导液培养hDPCs 14 d作为诱导组,以未诱导的hDPCs为对照组,实时定量PCR检测成牙本质相关基因：牙本质涎磷蛋白（dentin sialophosphoprotein,DSPP）、牙本质基质蛋白1（dentin matrix protein-1,DMP-1）、碱性磷酸酶（alkaline phosphatase,ALP）的表达;通过miRNA芯片筛选hDPCs向成牙本质细胞分化过程中差异表达的miRNAs,采用实时定量 PCR对结果进行验证;应用生物信息学软件进行靶基因预测,初步推测BMP-2诱导hDPCs分化过程中涉及的通路及功能。采用SPSS 18.0 软件包对数据进行统计学分析。结果实时定量PCR结果显示,各目的基因（DSPP、DMP-1、ALP）在诱导组的表达均高于对照组（P<0.05）。基因芯片结果显示,hDPCs成牙本质向诱导后,差异表达的miRNAs有36个,包括上调20个和下调16个。实时定量PCR对其中5个miRNAs的验证结果与芯片结果基本一致。生物信息学分析显示,基因的功能分布为37.8%与生物过程有关,29%与细胞成分有关,33%与分子功能有关,约有0.2%的基因未检测出具体功能和作用。结论在hDPCs经BMP-2诱导向成牙本质细胞分化过程中,存在miRNA表达谱变化,为进一步研究差异表达的miRNAs在hDPCs分化过程中的作用及其靶基因的生物学效应奠定了基础。

关键词: 牙髓细胞, BMP-2, miRNA, 牙本质细胞, 基因表达谱

Abstract: PURPOSE: To screen and verify the differentially expressed microRNAs (miRNAs) during the differentiation of human dental pulp cells (hDPCs) to odontoblasts induced by BMP-2. METHODS: The isolated hDPCs were cultured in vitro and induced by BMP-2. The levels of ALP, DMP-1 and DSPP were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). The potential characteristics of hDPCs were investigated by miRNA microarray and highly expressed miRNAs were selected with bio-information software for predicting target genes and their biological functions. Then the results were validated using qRT-PCR analysis for the selected miRNAs. Statistical analysis was performed using SPSS 18.0 software package. RESULTS: The expression of ALP, DSPP, and DMP-1 showed significantly higher levels in BMP-2 induced groups compared to the control group(P<0.05). A total of 36 miRNAs were changed (i.e. 20 miRNAs were up-regulated and 16 were down-regulated). The results of qRT-PCR were consistent with the microarray results. GO categories revealed that they were mainly associated with biological process(37.8%), cellular component (29%), molecular function(33%), while the function of other 0.2% genes remained unknown. CONCLUSIONS: This study identified differential expression of miRNAs in BMP-2-induced odontoblastic differentiation of hDPCs, thus contributing to further investigations of regulatory mechanisms and biological effect of target genes in BMP-2-induced odontoblastic differentiation of hDPCs.

Key words: Dental pulp cells, BMP-2, MiRNA, Odontoblasts, MiRNA Profile

中图分类号:

R781.3

包丽荣, 赵文青, 林田, 陆彦玲, 吴煜. BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化[J]. 上海口腔医学, 2017, 26(5): 476-483.

BAO Li-rong, ZHAO Wen-qing, LIN Tian, LU Yan-ling, WU Yu. miRNA profile of the human dental pulp cells during odontoblast differentiation induced by BMP-2[J]. Shanghai Journal of Stomatology, 2017, 26(5): 476-483.

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BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化

miRNA profile of the human dental pulp cells during odontoblast differentiation induced by BMP-2

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