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LncRNA H19 Promotes Odontoblastic Differentiation of Human Dental Pulp Stem Cells by Regulating miR-140-5p and BMP-2/FGF9
Qianzhou Jiang
Stomatology Hospital of Guangzhou Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2631-7527
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This work is licensed under a CC BY 4.0 License. Read Full License
Background: Increasing evidence has revealed that long non-coding RNAs (lncRNAs) exert critical roles in biological mineralization.As a critical process for dentin formation, odontoblastic differentiation is regulated by complex signaling networks. The present study aimed to investigate the biological role and regulatory mechanisms of lncRNA-H19 (H19) in regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs).
Methods: We performed lncRNA microarray assay to reveal the expression patterns of lncRNAs involved in odontoblastic differentiation. H19 was identified and verified as a critical factor by qRT-PCR. The gain- and loss-of-function studies were performed to investigate the biological role of H19 in regulating odontoblastic differentiation of hDPSCs in vitro and in vivo . Odontoblastic differentiation was evaluated through qRT-PCR, Western blot and Alizarin Red S staining. Bioinformatics analysis identified that H19 could directly interact with miR-140-5p, which was further verified by luciferase reporter assay. After overexpression of miR-140-5p in hDPSCs, odontoblastic differentiation was determined. Moreover, the potential target genes of miR-140-5p were investigated and the biological functions of BMP-2 and FGF9 in hDPSCs were verified. Co-transfection experiments were conducted to validate miR-140-5p was involved in H19-mediated odontoblastic differentiation in hDPSCs.
Results: The expression of H19 was significantly up-regulated in hDPSCs undergoing odontoblastic differentiation. Overexpression of H19 stimulated odontoblastic differentiation in vitro and in vivo , whereas down-regulation of H19 revealed the opposite effect. H19 binds directly to miR-140-5p and overexpression of miR-140-5p inhibited odontoblastic differentiation of hDPSCs. H19 acted as a miR-140-5p sponge, resulting in regulated the expression of BMP-2 and FGF9. Overexpression of H19 abrogated the inhibitory effect of miR-140-5p on odontoblastic differentiation.
Conclusion: Our data revealed that H19 plays a positive regulatory role in odontoblastic differentiation of hDPSCs through miR-140-5p/BMP-2/FGF9 axis, suggesting that H19 may be a stimulatory regulator of odontogenesis.
Keywords
long non-coding RNA, H19, human dental pulp stem cells (hDPSCs), dentinogenesis, odontoblastic differentiation, miR-140-5p
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CURRENT STATUS: Under Review
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Received 24 Apr, 2020
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On 14 Apr, 2020
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On 13 Apr, 2020
Review #1 received
Received 13 Apr, 2020
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On 07 Apr, 2020
Reviewers invited
Invitations sent on 07 Apr, 2020
Submission checks complete
On 06 Apr, 2020
Editor invited
On 06 Apr, 2020
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Review #1 received
Received 10 Mar, 2020
Review #2 received
Received 10 Mar, 2020
Editorial decision: Major Revision
On 10 Mar, 2020
Reviewer #2 agreed
On 05 Mar, 2020
Reviewers invited
Invitations sent on 04 Mar, 2020
Reviewer #1 agreed
On 04 Mar, 2020
Editor assigned
On 03 Mar, 2020
Editor invited
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This preprint is under consideration at Stem Cell Research & Therapy. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
LncRNA H19 Promotes Odontoblastic Differentiation of Human Dental Pulp Stem Cells by Regulating miR-140-5p and BMP-2/FGF9
Qianzhou Jiang
Stomatology Hospital of Guangzhou Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2631-7527
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 2
Posted 14 Apr, 2020
No community comments so far
Review #2 received
Received 24 Apr, 2020
Reviewer #2 agreed
On 14 Apr, 2020
Reviewer #1 agreed
On 13 Apr, 2020
Review #1 received
Received 13 Apr, 2020
Editor assigned
On 07 Apr, 2020
Reviewers invited
Invitations sent on 07 Apr, 2020
Submission checks complete
On 06 Apr, 2020
Editor invited
On 06 Apr, 2020
No comments provided
Review #1 received
Received 10 Mar, 2020
Review #2 received
Received 10 Mar, 2020
Editorial decision: Major Revision
On 10 Mar, 2020
Reviewer #2 agreed
On 05 Mar, 2020
Reviewers invited
Invitations sent on 04 Mar, 2020
Reviewer #1 agreed
On 04 Mar, 2020
Editor assigned
On 03 Mar, 2020
Editor invited
On 02 Mar, 2020
Submission checks complete
On 02 Mar, 2020
First submitted
On 29 Feb, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Increasing evidence has revealed that long non-coding RNAs (lncRNAs) exert critical roles in biological mineralization.As a critical process for dentin formation, odontoblastic differentiation is regulated by complex signaling networks. The present study aimed to investigate the biological role and regulatory mechanisms of lncRNA-H19 (H19) in regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs).
Methods: We performed lncRNA microarray assay to reveal the expression patterns of lncRNAs involved in odontoblastic differentiation. H19 was identified and verified as a critical factor by qRT-PCR. The gain- and loss-of-function studies were performed to investigate the biological role of H19 in regulating odontoblastic differentiation of hDPSCs in vitro and in vivo . Odontoblastic differentiation was evaluated through qRT-PCR, Western blot and Alizarin Red S staining. Bioinformatics analysis identified that H19 could directly interact with miR-140-5p, which was further verified by luciferase reporter assay. After overexpression of miR-140-5p in hDPSCs, odontoblastic differentiation was determined. Moreover, the potential target genes of miR-140-5p were investigated and the biological functions of BMP-2 and FGF9 in hDPSCs were verified. Co-transfection experiments were conducted to validate miR-140-5p was involved in H19-mediated odontoblastic differentiation in hDPSCs.
Results: The expression of H19 was significantly up-regulated in hDPSCs undergoing odontoblastic differentiation. Overexpression of H19 stimulated odontoblastic differentiation in vitro and in vivo , whereas down-regulation of H19 revealed the opposite effect. H19 binds directly to miR-140-5p and overexpression of miR-140-5p inhibited odontoblastic differentiation of hDPSCs. H19 acted as a miR-140-5p sponge, resulting in regulated the expression of BMP-2 and FGF9. Overexpression of H19 abrogated the inhibitory effect of miR-140-5p on odontoblastic differentiation.
Conclusion: Our data revealed that H19 plays a positive regulatory role in odontoblastic differentiation of hDPSCs through miR-140-5p/BMP-2/FGF9 axis, suggesting that H19 may be a stimulatory regulator of odontogenesis.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6