MiR-34a在牙周膜细胞成骨向分化中的作用

doi:10.13701/j.cnki.kqyxyj.2017.09.005

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (9): 928-932.DOI: 10.13701/j.cnki.kqyxyj.2017.09.005

MiR-34a在牙周膜细胞成骨向分化中的作用

程孟文¹，周毅^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. 武汉大学口腔医院修复科湖北武汉 430079

收稿日期:2017-01-20 出版日期:2017-09-20 发布日期:2017-09-27
通讯作者: 周毅,E-mail: dryizhou@163.com
作者简介:程孟文(1995~ ),女,安徽阜阳人,硕士在读,主要从事口腔修复学研究工作。
基金资助:
国家自然科学基金(编号:81200812)武汉市应用基础研究计划项目(编号:2016060101010043)

Role of Micro-34a in Induced-mineralization of Human Periodontal Ligament Cells

CHENG Meng-wen¹, ZHOU Yi^1,2*

1. The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China;
2. Department of Prosthodontics, Hospital of Stomatology, Wuhan University, Wuhan 430079, China.

Received:2017-01-20 Online:2017-09-20 Published:2017-09-27

摘要/Abstract

摘要： 目的:探讨微小RNA-34a在牙周膜细胞成骨向分化中的作用。方法:体外分离培养人牙周膜细胞(hPDLCs),取第3~6代用于实验。首先,对hPDLCs进行成骨诱导液处理,在3、7、14 d后利用实时荧光定量PCR(qRT-PCR)检测miR-34a基因的表达变化。然后,构建慢病毒载体pCDH-pre-miR-34a和pCDH,将慢病毒载体感染hPDLCs,构建pre-miR-34a过表达细胞模型(hPDLCs/34a)和空载体对照细胞模型(hPDLCs/pCDH),并诱导hPDLCs成骨分化3、7、14和21 d。分析成骨标志基因碱性磷酸酶(ALP)、Runt相关转录因子2(Runx2)、骨钙素(OCN)及骨涎蛋白(BSP)表达变化,ALP活性及钙化结节形成的茜素红染色情况。结果:hPDLCs经成骨分化诱导后, miR-34a基因表达在第3天开始明显升高,并呈逐渐增高趋势,差异均有统计学意义(P<0.001)。与hPDLCs/pCDH组相比,hPDLCs/34a组的ALP活性和茜素红染色均有明显减弱。qRT-PCR结果显示:hPDLCs/34a组的ALP表达量在成骨诱导7 d时较hPDLCs/pCDH组降低(P<0.05);Runx2、OCN及BSP表达量的差异在各时间点基本无统计学意义。结论:在体外条件下,miR-34a抑制人牙周膜细胞成骨向分化。

关键词: 人牙周膜细胞, 成骨分化, 微小RNA-34a, 实时荧光定量PCR

Abstract: Objective: To investigate the effect of miR-34a on the osteogenic differentiation of human periodontal ligament cells (hPDLCs). Methods: HPDLCs were obtained through in vitro culture combined with enzyme digestion method and tissue pieces culture method. First, the cells were continuously induced by mineralization induction medium for 3,7, and 14 days. The expression of miR-34a was detected by quantitative real-time PCR (qRT-PCR). Second, hPDLCs were transfected with lentiviral vector containing pre-miR-34a or control to build the miR-34a overexpression model or control model (hPDLCs/34a or hPDLCs/pCDH) and induced by mineralization induction medium for 3, 7, 14, and 21 days. Quantification of osteogenensis-related genes (ALP, Runx2, OCN and BSP) expression, alkaline phosphatase (ALP) activity detection and Alizarin red staining were used to assess the influence of miR-34a on the osteogenic differentiation of hPDLCs. Results: The expression of miR-34a significantly increased in a time-dependent manner (P<0.001) during the osteogenic differentiation. The mineralization nodule formation and ALP activity were all attenuated by miR-34a. ALP gene expression detected by qRT-PCR was reduced after 7 days' induction in hPDLCs/34a. Runx2, OCN and BSP gene expression showed no significant

基金项目国家自然科学基金(编号:81200812)
武汉市应用基础研究计划项目(编号:2016060101010043)
作者简介程孟文(1995~ ),女,安徽阜阳人,硕士在读,主要从事口腔修复学研究工作。
*通讯作者周毅,E-mail: dryizhou@163.com
differences. Conclusion: MiR-34a may have a negative effect on the osteogenic differentiation of hPDLCs.

Key words: Human periodontal ligament cells , Osteogenic differentiation, MicroRNA-34a, Quantitative real-time PCR

中图分类号:

R329.2

程孟文，周毅. MiR-34a在牙周膜细胞成骨向分化中的作用[J]. 口腔医学研究, 2017, 33(9): 928-932.

CHENG Meng-wen, ZHOU Yi. Role of Micro-34a in Induced-mineralization of Human Periodontal Ligament Cells[J]. Journal of Oral Science Research, 2017, 33(9): 928-932.

参考文献

[1] Agostini M, Knight RA. miR-34: from bench to bedside [J]. Oncotarget, 2014, 5(4)∶872-881
[2] Li X J, Ren ZJ, Tang JH. MicroRNA-34a: a potential therapeutic target in human cancer [J]. Cell Death Dis, 2014, 5∶e1327
[3] Krzeszinski JY, Wei W, Huynh H, et al. miR-34a blocks osteoporosis and bone metastasis by inhibiting osteoclastogenesis and Tgif2 [J]. Nature, 2014, 512(7515)∶431-435
[4] Chen L, Holmstrom K, Qiu W, et al. MicroRNA-34a inhibits osteoblast differentiation and in vivo bone formation of human stromal stem cells [J]. Stem Cells, 2014, 32(4)∶902-912
[5] Fan C, Jia L, Zheng Y, et al. MiR-34a promotes osteogenic differentiation of human adipose-derived stem cells via the RBP2/NOTCH1/CYCLIN D1 coregulatory network [J]. Stem Cell Reports, 2016, 7(2)∶236-248
[6] He Y, de Castro LF, Shin MH, et al. p53 loss increases the osteogenic differentiation of bone marrow stromal cells [J]. Stem Cells, 2015, 33(4)∶1304-1319
[7] 廖海清,曹正国.经典Wnt信号通路在牙周膜细胞成骨分化过程中的调控[J].口腔医学研究,2016,32(2)∶224-227
[8] Lee YH, Na H S, Jeong SY, et al. Comparison of inflammatory microRNA expression in healthy and periodontitis tissues [J]. Biocell, 2011, 35(2)∶43-49
[9] Ogata Y, Matsui S, Kato A, et al. MicroRNA expression in inflamed and noninflamed gingival tissues from Japanese patients [J]. J Oral Sci, 2014, 56(4)∶253-260
[10] Jiang P, Liu R, Zheng Y, et al. MiR-34a inhibits lipopolysaccharide-induced inflammatory response through targeting Notch1 in murine macrophages [J]. Exp Cell Res, 2012, 318(10)∶1175-1184
[11] Li Y, Li S Q, Gao YM, et al. Crucial role of Notch signaling in osteogenic differentiation of periodontal ligament stem cells in osteoporotic rats [J]. Cell Biol Int, 2014, 38(6)∶729-736
[12] Fan W, Fang R, Wu X, et al. Shear-sensitive microRNA-34a modulates flow-dependent regulation of endothelial inflammation [J]. J Cell Sci, 2015, 128(1)∶70-80
[13] Badi I, Burba I, Ruggeri C, et al. MicroRNA-34a induces vascular smooth muscle cells senescence by SIRT1 downregulation and promotes the expression of age-associated pro-inflammatory secretory factors [J]. J Gerontol A Biol Sci Med Sci, 2015, 70(11)∶1304-1311
[14] Lee YM, Shin SI, Shin KS, et al. The role of sirtuin 1 in osteoblastic differentiation in human periodontal ligament cells [J]. J Periodontal Res, 2011, 46(6)∶712-721
[15] Zhang QB, Cao W, Liu YR, et al. Effects of Sirtuin 1 on the proliferation and osteoblastic differentiation of periodontal ligament stem cells and stem cells from apical papilla [J]. Genet Mol Res, 2016, 15(1)∶

MiR-34a在牙周膜细胞成骨向分化中的作用

Role of Micro-34a in Induced-mineralization of Human Periodontal Ligament Cells

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