Abstract
Accumulating research continues to highlight the notable role of microRNAs (miRs) and long non-coding RNAs (lncRNAs) as important regulators in the process of human dental pulp stem cell (hDPSCs) differentiation. The current study aimed to investigate the novel regulatory circuitry of lncRNA metastasis–associated lung adenocarcinoma transcript 1 (MALAT1)/miR-140-5p/G protein–coupled receptor (GPCR)–kinase 2 interacting protein 2 (GIT2) on the odontogenic differentiation of hDPSCs. In hDPSCs, miR-140-5p was downregulated during the odontogenic differentiation, which was verified to directly target GIT2. RNA crosstalk determined by dual-luciferase reporter and RNA pull-down assays revealed that MALAT1 could bind to miR-140-5p to upregulate the expression of GIT2. After that, the levels of MALAT1, miR-140-5p, and GIT2 in hDPSCs were up- or downregulated by exogenous transfection or lentivirus infection in order to investigate their effects on the differentiation of hDPSCs. It was observed that elevation of miR-140-5p or knockdown of GIT2 resulted in inhibited alkaline phosphatase (ALP) activity, expression of dentin sialophosphoprotein (DSPP), dentin matrix-protein-1 (DMP-1), and distal-less homeobox 3 (DLX3) as well as positive expression of desmoplakin (DSP) protein. The promotive effects of MALAT1 on odontogenic differentiation were diminished by restoration of miR-140-5p or inhibition of GIT2. Taken together, this study provides valuable evidence suggesting MALAT1 as a potential contributor to the odontogenic differentiation of hDPSCs.
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Abbreviations
- miRs:
-
microRNAs
- lncRNAs:
-
long non-coding RNAs
- hDPSCs:
-
human dental pulp stem cell
- MALAT1:
-
metastasis-associated lung adenocarcinoma transcript 1
- GPCR:
-
G protein–coupled receptor
- ALP:
-
alkaline phosphatase
- DSPP:
-
dentin sialophosphoprotein
- DMP-1:
-
dentin matrix-protein-1
- DLX3:
-
distal-less homeobox 3
- DSP:
-
desmoplakin
- MSC:
-
mesenchymal stem cell
- miR-508-5p:
-
microRNA-508-5p
- FBS:
-
fetal bovine serum
- PBS:
-
phosphate-buffered saline
- FITC:
-
fluorescein isothiocyanate
- OM:
-
odontogenic induction medium
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- HRP:
-
horseradish peroxidase
- ECL:
-
enhanced chemiluminescence
- 3’UTR:
-
3′-untranslated region
- wt:
-
wild type
- RT-qPCR:
-
reverse transcription quantitative PCR
- ANOVA:
-
analysis of variance
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We would like to give our sincere appreciation to our colleagues for their valuable efforts on this paper.
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The authors received support from the Science and Technology Plan of Liaoning Province, Grant/Award Number: 2013225090.
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All participants signed written informed consents. The current study was conducted under the approval of the Ethics Committee of School and Hospital of Stomatology, China Medical University and carried out in accordance with the Helsinki declaration.
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Bao, M., Liu, G., Song, J. et al. Long non-coding RNA MALAT1 promotes odontogenic differentiation of human dental pulp stem cells by impairing microRNA-140-5p-dependent downregulation of GIT2. Cell Tissue Res 382, 487–498 (2020). https://doi.org/10.1007/s00441-020-03246-1
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DOI: https://doi.org/10.1007/s00441-020-03246-1