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lncRNA DANCR suppresses odontoblast-like differentiation of human dental pulp cells by inhibiting wnt/β-catenin pathway

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Abstract

Long noncoding RNAs (lncRNAs) have recently emerged as an important class of regulatory molecules in diverse biological processes, although lncRNA involvement in the odontoblast-like differentiation of human dental pulp cells (hDPCs) is poorly understood. We investigate the expression of lncRNAs in this differentiation and explore their underlying role and the involved mechanism. Integrated comparative lncRNA microarray profiling was used to examine lncRNA expression during this differentiation. The differential expression of lncRNAs was validated by quantitative real-time reverse transcription plus the polymerase chain reaction. Differential lncRNA overexpression was performed with an adenoviral vector and the role and mechanism was then investigated in odontoblast-like differentiation. We identified 139 differentially expressed lncRNAs during this differentiation. Among them, five lncRNAs differentially expressed in microarray analysis were validated. Notably, lncRNA DANCR expression was significantly downregulated during hDPC differentiation to odontoblast-like cells in a time-dependent manner. Moreover, lncRNA DANCR overexpression blocked mineralized nodule formation and the expression of DSPP and DMP-1 in hDPCs after 14 days of odontogenic induction. Importantly, the upregulation of DANCR significantly decreased the expression levels of p-GSK-3β and β-catenin expression indicating that lncRNA DANCR can inhibit the activation of the Wnt/β-catenin signal pathway during the odontoblast-like differentiation of hDPCs. Thus, the modulation of Wnt/β-catenin signaling by lncRNA DANCR represents a potential therapeutic option for reparative dentin formation and regenerative endodontics.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (grant no. 81070829; 81271124).

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Authors and Affiliations

  1. Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University and Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong, 510055, People’s Republic of China

    Lingling Chen, Zhi Song, Shuheng Huang, Runfu Wang, Wei Qin, Jia Guo & Zhengmei Lin

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  1. Lingling Chen
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  2. Zhi Song
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  3. Shuheng Huang
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  4. Runfu Wang
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  5. Wei Qin
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  6. Jia Guo
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  7. Zhengmei Lin
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Correspondence to Zhengmei Lin.

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The authors declare that they have no conflicting financial interests.

Additional information

Lingling Chen and Zhi Song contributed equally to this work.

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Electronic Supplementary Material, Fig. S1

Characteristics of hDPCs during odontoblast-like differentiation. a Morphology of cells at the third passage. b Alkaline phosphatase activity (ALP) at various times in odontogenic-inductive medium (OM). c Appearance of hDPCs after 14 days in non-differentiation medium. d hDPCs after 14 days in OM and with alizarin red staining (ARS) for mineralization (compare with control cells in c). e Total area of mineralized nodules was quantified. f Transcript levels of odontogenic-related genes expressed relative to glyceraldehyde-3-phosphate dehydrogenase (GADPH). g Levels of odontogenic-related proteins. h, i Quantitative analysis of ontogenic-related proteins by Western blot (DSPP dentin sialophosphoprotein, DMP-1 dentin matrix protein-1). Data are expressed as means ± SD. *P< 0.05 compared with cells cultured for 0 days; # P< 0.05 compared with undifferentiated controls. Bar 200 μm. (GIF 118 kb)

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Electronic Supplementary Material, Table S1

Differentially expressed lncRNAs in odontoblast-like differentiation of hDPCs (more than two-fold; P<0.05). (XLSX 42 kb)

Electronic Supplementary Material, Table S2

Differentially expressed mRNAs in odontoblast-like differentiation of hDPCs (more than two-fold; P<0.05). (XLSX 57 kb)

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Chen, L., Song, Z., Huang, S. et al. lncRNA DANCR suppresses odontoblast-like differentiation of human dental pulp cells by inhibiting wnt/β-catenin pathway. Cell Tissue Res 364, 309–318 (2016). https://doi.org/10.1007/s00441-015-2333-2

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  • DOI: https://doi.org/10.1007/s00441-015-2333-2

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