RUNX3 derived hsa_circ_0005752 accelerates the osteogenic differentiation of adipose-derived stem cells via the miR-496/MDM2-p53 pathway

Autor: Yifan Huan, Ming Wang, Jing Li, Guohua Lv, Xiyang Li
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Adipose-derived stem cells
Medicine (General)
3′ UTR
3′ untranslated region
ECL
enhanced chemiluminescence
BM-MSCs
Bone Marrow-Mesenchymal Stem Cells
H&E staining
Hematoxylin and Eosin staining
Osteogenic differentiation
Transcriptional regulation
Gene knockdown
medicine.diagnostic_test
biology
microRNA
Chemistry
qRT-PCR
quantitative real-time polymerase chain reaction
RIP
RNA immunoprecipitation
LPAR1
lysophosphatidic acid receptor 1
ARS
Alizarin Red Staining
Cell biology
RUNX2
ChIP
chromatin immunoprecipitation
OM
osteogenic (differentiation) medium
Mdm2
Alkaline phosphatase
Original Article
BMP2
Bone morphogenetic protein 2
circRNAs
Circular RNAs
ADSCs
adipose-derived stem cells
Circular RNAs
RUNX3
Biomedical Engineering
miRNAs
microRNA
Runx3
RUNX Family Transcription Factor 3
Biomaterials
R5-920
Western blot
MDM2
Osx
osterix
medicine
SDS-PAGE
polyacrylamide gel electrophoresis
Messenger RNA
QH573-671
MDM2
murine double minute 2
ALP
alkaline phosphatase
digestive system diseases
BCA
bicinchoninic acid
OCN
osteocalcin
PMSF
phenylmethylsulfonyl fluoride
biology.protein
UC-MSCs
Umbilical Cord-Mesenchymal Stem Cells
Cytology
Runx2
Runt-related transcription factor 2
Developmental Biology
Zdroj: Regenerative Therapy
Regenerative Therapy, Vol 18, Iss, Pp 430-440 (2021)
ISSN: 2352-3204
Popis: Background Circular RNAs (circRNAs) are non-coding RNAs that play a pivotal role in bone diseases. RUNX3 was an essential transcriptional regulator during osteogenesis. However, it is unknown whether RUNX3 regulates hsa_circ_0005752 during osteogenic differentiation. Methods The levels of hsa_circ_0005752 and RUNX3 were measured by qRT-PCR after osteogenic differentiation of ADSCs. The osteogenic differentiation was analyzed by Alkaline phosphatase (ALP) staining and Alizarin red staining (ARS). qRT-PCR and western blot were used to assess the expressions of osteogenic differentiation-related molecules. RNA pull-down, RIP, and luciferase reporter assays determine the interactions between miR-496 and hsa_circ_0005752 or MDM2 mRNA. CHIP-PCR analyzed the interaction between RUNX3 and LPAR1. Finally, the potential roles of RUNX3 were investigated during osteogenic differentiation with or without hsa_circ_0005752 knockdown. Results Hsa_circ_0005752 and RUNX3 were significantly increased, and miR-496 was remarkably decreased in ADSCs after osteogenic differentiation. Hsa_circ_0005752 could promote osteogenic differentiation, as shown by enhancing ALP and ARS staining intensity. Hsa_circ_0005752 enhanced the expressions of Runx2, ALP, Osx, and OCN. Furthermore, hsa_circ_0005752 directly targeted miR-496, which can directly bind to MDM2. RUNX3 bound to the LPAR1 promoter and enhanced hsa_circ_0005752 expressions. Moreover, the enhanced expression of hsa_circ_0005752 by RUNX3 could promote osteogenic differentiation, whereas knockdown of hsa_circ_0005752 partially antagonized the effects of RUNX3. Conclusion Our study demonstrated that RUNX3 promoted osteogenic differentiation via regulating the hsa_circ_0005752/miR-496/MDM2 axis and thus provided a new therapeutic strategy for osteoporosis.
Databáze: OpenAIRE
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