» Articles » PMID: 25817070

MiR-20a Regulates Adipocyte Differentiation by Targeting Lysine-specific Demethylase 6b and Transforming Growth Factor-β Signaling

Overview
Specialty Endocrinology
Date 2015 Mar 31
PMID 25817070
Citations 18
Authors
Affiliations
Soon will be listed here.
Abstract

Background: Several types of microRNAs (miRNAs) have recently been defined as important regulators in adipocyte differentiation, the role of other miRNAs in the processes and the mechanisms involved remain to be explored.

Methods: miR-20a expression was quantified in primary cultured marrow stromal cells and adipogenic cell lines after adipogenic treatment. Effects of miR-20a on adipocyte differentiation were studied following supplementing or depleting miR-20a in murine 3T3-L1 preadipocytes, ST2 stromal cells and C3H10T1/2 mesenchymal cells. Bioinformatics prediction of miRNA targets was performed, and potential targets of miR-20a were verified by using dual luciferase activity assays. Gain-of-function and loss-of-function studies were performed to examine the effects of the target genes on adipocyte differentiation.

Results: miR-20a was induced in primary cultured marrow stromal cells and established adipogenic lines after adipogenic treatment. Supplementing miR-20a activity suppressed the growth of 3T3-L1 preadipocytes and induced 3T3-L1, ST2 and C3H10T1/2 cells to differentiate into mature adipocytes, along with the induction of adipocyte-specific transcription factors peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα), C/EBPβ and the marker gene adipocyte protein 2 (aP2). Conversely, inhibition of the endogenous miR-20a repressed 3T3-L1, ST2 and C3H10T1/2 cells to fully differentiate. Transforming growth factor-β receptor II (Tgfbr2) and lysine-specific demethylase 6b (Kdm6b) were shown to be direct targets of miR-20a. Supplementing miR-20a activity in ST2 reduced levels of KDM6B and TGFBR2 proteins, while suppression of endogenous miR-20a increased KDM6B and TGFBR2. While TGF-β signaling is a well-documented inhibitor of adipogenesis, the effects of Kdm6b on adipocyte formation need to be clarified. We demonstrated that overexpression of Kdm6b inhibited, while knockdown of Kdm6b promoted the differentiation of the ST2 cells into mature adipocytes.

Conclusion: The present work provides evidence that mouse miR-20a promotes adipocyte progenitor cells to differentiate and this function may depend upon its inhibitory effects on Kdm6b and TGF-β signaling.

Citing Articles

The Therapeutic Potential of MicroRNA-21 in the Treatment of Spinal Cord Injury.

Hasan A, Ardizzone A, Giosa D, Scuderi S, Calcaterra E, Esposito E Curr Issues Mol Biol. 2025; 47(2).

PMID: 39996791 PMC: 11854632. DOI: 10.3390/cimb47020070.


Mechanism of MiR-145a-3p/Runx2 pathway in dexamethasone impairment of MC3T3-E1 osteogenic capacity in mice.

Wu H, Liao X, Wu T, Xie B, Ding S, Chen Y PLoS One. 2024; 19(11):e0309951.

PMID: 39561180 PMC: 11575826. DOI: 10.1371/journal.pone.0309951.


Declined adipogenic potential of senescent MSCs due to shift in insulin signaling and altered exosome cargo.

Voynova E, Kulebyakin K, Grigorieva O, Novoseletskaya E, Basalova N, Alexandrushkina N Front Cell Dev Biol. 2022; 10:1050489.

PMID: 36467400 PMC: 9714334. DOI: 10.3389/fcell.2022.1050489.


Regulatory of miRNAs in tri-lineage differentiation of C3H10T1/2.

Huang W, Wu X, Xiang S, Qiao M, Li H, Zhu Y Stem Cell Res Ther. 2022; 13(1):521.

PMID: 36414991 PMC: 9682817. DOI: 10.1186/s13287-022-03205-3.


Profiling Secreted miRNA Biomarkers of Chemical-Induced Neurodegeneration in Human iPSC-Derived Neurons.

You D, Cohen J, Pustovalova O, Lewis L, Shen L Toxicol Sci. 2022; 186(2):221-241.

PMID: 35134991 PMC: 8963304. DOI: 10.1093/toxsci/kfac011.


References
1.
Freytag S, Paielli D, Gilbert J . Ectopic expression of the CCAAT/enhancer-binding protein alpha promotes the adipogenic program in a variety of mouse fibroblastic cells. Genes Dev. 1994; 8(14):1654-63. DOI: 10.1101/gad.8.14.1654. View

2.
Chen C, Ridzon D, Broomer A, Zhou Z, Lee D, Nguyen J . Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005; 33(20):e179. PMC: 1292995. DOI: 10.1093/nar/gni178. View

3.
Tsang W, Kwok T . The miR-18a* microRNA functions as a potential tumor suppressor by targeting on K-Ras. Carcinogenesis. 2009; 30(6):953-9. DOI: 10.1093/carcin/bgp094. View

4.
Byun M, Jeong H, Bae S, Kim A, Hwang E, Hong J . TAZ is required for the osteogenic and anti-adipogenic activities of kaempferol. Bone. 2011; 50(1):364-72. DOI: 10.1016/j.bone.2011.10.035. View

5.
Li H, Li T, Wang S, Wei J, Fan J, Li J . miR-17-5p and miR-106a are involved in the balance between osteogenic and adipogenic differentiation of adipose-derived mesenchymal stem cells. Stem Cell Res. 2013; 10(3):313-24. DOI: 10.1016/j.scr.2012.11.007. View