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.

参考文献

[1] Fabian MR,Sonenberg N. The mechanics of miRNA-mediated gene silencing: a look under the hood of miRISC [J]. Nat Struct Mol Biol, 2012, 19(6): 586-593.
[2] Guo L, Zhao RC, Wu Y. The role of microRNAs in self-renewal and differentiation of mesenchymal stem cells[J]. Exp Hematol, 2011, 39(6): 608-616.
[3] Qin W, Liu P, Zhang R, et al. JNK MAPK is involved in BMP-2-induced odontoblastic differentiation of human dental pulp cells [J]. Connect Tissue Res, 2014, 55(3): 217-224.
[4] Yang W, Harris MA, Cui Y, et al. Bmp2 is required for odontoblast differentiation and pulp vasculogenesis[J]. J Dent Res, 2012, 91(1): 58-64.
[5] Zhang W, Zhang X, Ling J, et al. Osteo-/odontogenic differentiation of BMP2 and VEGF gene-co-transfected human stem cells from apical papilla[J]. Mol Med Rep, 2016, 13(5): 3747-3754.
[6] Wu L, Wang F, Donly KJ, et al. Establishment of immortalized mouse BMP2 knock-out dental papilla mesenchymal cells necessary for study of odontoblastic differentiation and odontogenesis [J]. J Cell Physiol, 2015, 230(11): 2588-2595.
[7] 韩跃华, 刘文忠, 史耀舟, 等. 幽门螺杆菌基因组表达谱芯片的研制 [J]. 微生物学通报, 2007, 34(1): 6-9.
[8] 高冬梅. 新疆维吾尔族高危HPV感染宫颈鳞癌组织中miRNA的初步研究 [D]. 乌鲁木齐: 新疆医科大学, 2014.
[9] 李刚, 容敏华, 钟艳平, 等. EGCG诱导人肝癌SMMC-7721细胞早期凋亡及基因和蛋白谱的变化研究 [J]. 中国药理学通报, 2009, 25(11): 1482-1486.
[10] Tasli PN, Aydin S, Yalvac ME, et al. Bmp 2 and bmp 7 induce odonto- and osteogenesis of human tooth germ stem cells [J]. Appl Biochem Biotechnol, 2014, 172(6): 3016-3025.
[11] Wang Y, Li L, Zheng Y, et al. BMP activity is required for tooth development from the lamina to bud stage [J]. J Dent Res, 2012, 91(7): 690-695.

[1]	徐方芳, 蒋备战. DKK1对脂多糖感染人牙髓细胞生物学行为的影响[J]. 上海口腔医学, 2021, 30(4): 355-359.
[2]	代书林, 王旭霞, 乜福娇, 崔群, 韩媛媛, 张君. 异普黄酮对大鼠正畸牙复发过程中牙周组织改建的影响[J]. 上海口腔医学, 2021, 30(1): 23-27.
[3]	李静, 张艺, 王金华. 骨形态发生蛋白-2、-4、-6、-7和-9差异性介导永生化成牙本质细胞成骨分化[J]. 上海口腔医学, 2020, 29(3): 242-249.
[4]	刘青, 陈菲, 卢成焱, 李静, 李晓乐, 张君. 槲皮素在大鼠上颌快速扩弓时对腭中缝骨改建的影响[J]. 上海口腔医学, 2020, 29(1): 1-6.
[5]	曹颖, 彭伟伟, 王璇, 伍甜甜, 杜嵘, 朱亚琴. 重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响[J]. 上海口腔医学, 2020, 29(1): 7-12.
[6]	彭伟伟, 戴兆威, 曹颖, 韩俊力, 朱亚琴. 血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响[J]. 上海口腔医学, 2019, 28(1): 1-5.
[7]	林田, 赵文青, 陆彦玲, 刘玉莹, 包丽荣, 吴煜. 人脐带间充质干细胞与牙髓细胞体外共培养对细胞生物学的影响[J]. 上海口腔医学, 2018, 27(4): 365-369.
[8]	李莉芬, 文扬, 江龙, 朱亚琴. 衣霉素诱导牙髓细胞内质网应激模型的建立[J]. 上海口腔医学, 2018, 27(2): 135-138.
[9]	杜建新, 胡玮. Krüpple样因子5在成牙本质细胞中的表达及临床意义[J]. 上海口腔医学, 2017, 26(6): 582-585.
[10]	徐超, 王旭霞, 王英姿, 杨盼盼, 赵姝亚, 张君. 雷奈酸锶对大鼠快速扩弓过程中腭中缝BMP-2表达的影响[J]. 上海口腔医学, 2016, 25(6): 647-651.
[11]	韦晓玲, 刘道杨, 潘杰, 陆珮珺, 赵君. 冻干对BMP-2基因重组慢病毒载体生物学效应的影响[J]. 上海口腔医学, 2016, 25(5): 522-526.
[12]	郝文秀, 张云涛, 张雅杰. 桂皮醛对成骨细胞成骨功能及BMP-2 mRNA表达的影响[J]. 上海口腔医学, 2016, 25(3): 271-274.
[13]	梁雪艺，胡靖宇，周刚. miR-155、miR-146a在口腔扁平苔藓患者外周血和组织中的变化与相关性研究[J]. 上海口腔医学, 2015, 24(6): 729-734.
[14]	何璇,韦维,陈文霞. 富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响[J]. 上海口腔医学, 2015, 24(3): 263-268.
[15]	王艳丽, 潘克清, 孙艳, 邓婧. 脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的影响[J]. 上海口腔医学, 2014, 23(4): 431-435.

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

PDF (PC)

可视化

被引次数

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

本文评价