miR‑204-5p promotes the adipogenic differentiation of human adipose-derived mesenchymal stem cells by modulating DVL3 expression and suppressing Wnt/β-catenin signaling

He,Honghui; Chen,Ke; Wang,Fang; Zhao,Liling; Wan,Xinxing; Wang,Linghao; Mo,Zhaohui

doi:10.3892/ijmm.2015.2160

miR‑204-5p promotes the adipogenic differentiation of human adipose-derived mesenchymal stem cells by modulating DVL3 expression and suppressing Wnt/β-catenin signaling

Authors:
- Honghui He
- Ke Chen
- Fang Wang
- Liling Zhao
- Xinxing Wan
- Linghao Wang
- Zhaohui Mo
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Affiliations: Department of Endocrinology, Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, P.R. China
Published online on: March 31, 2015 https://doi.org/10.3892/ijmm.2015.2160
Pages: 1587-1595
Copyright: © He et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

MicroRNAs (miRNAs or miRs) play an important regulatory role during adipogenesis, and have been studied extensively in this regard. Specifically, the switch between the differentiation of mesenchymal stem cells (MSCs) towards adipogenic vs. osteogenic lineages is regulated by miR‑204 which controls the expression of Runx2. However, the association between miR‑204-5p and the Wnt/β-catenin signaling pathway during adipogenesis has not yet been clarified. In the present study, we demonstrate that miR‑204-5p regulates the in vitro adipogenesis of human adipose-derived mesenchymal stem cells (hADSCs). The level of miR‑204-5p was shown to be gradually upregulated during adipocytic differentiation, together with the mRNA expression of the critical adipogenic transcription factors, cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT) enhancer binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ), and the mature adipogenic marker, fatty acid binding protein 4 (FABP4). We further demonstrate that while the overexpression of miR‑204-5p promotes adipogenesis, its knockdown causes the inhibition of this process. We then used bioinformatics tools and luciferase reporter assay to establish that dishevelled segment polarity protein 3 (DVL3), a key regulator of the Wnt/β-catenin signaling pathway, is a direct target of miR‑204-5p. In addition, the overexpression of DVL3 led to an increase in β-catenin and cyclin D1 (CCND1) expression and, by contrast, the knockdown of DVL3 led to a decrease in the expression of β-catenin and CCND1. The knockdown of DVL3 significantly promoted adipogenesis. Finally, we demonstrated that the overexpression of miR‑204-5p induced the downregulation of β-catenin and the canonical Wnt target gene, CCND1, in mature adipoctyes, while its knockdown led to their upregulation. Taken together, our data suggest that miR‑204-5p regulates adipogenesis by controlling DVL3 expression and subsequently inhibiting the activation of the Wnt/β-catenin signaling pathway.

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1	Kanuri G and Bergheim I: In vitro and in vivo models of non-alcoholic fatty liver disease (NAFLD). Int J Mol Sci. 14:11963–11980. 2013. View Article : Google Scholar : PubMed/NCBI
2	Sung YY, Yoon T, Yang WK, Moon BC and Kim HK: Anti-obesity effects of Actinidia polygama extract in mice with high-fat diet-induced obesity. Mol Med Rep. 7:396–400. 2013.
3	Ma J, Jiang Z, He S, Liu Y, Chen L, Long K, Jin L, Jiang A, Zhu L, Wang J, Li M and Li X: Intrinsic features in microRNA transcriptomes link porcine visceral rather than subcutaneous adipose tissues tometabolic risk. PLoS One. 8:e800412013. View Article : Google Scholar
4	Song G, Xu G, Ji C, Shi C, Shen Y, Chen L, Zhu L, Yang L, Zhao Y and Guo X: The role of microRNA-26b in human adipocyte differentiation and proliferation. Gene. 10:481–487. 2014. View Article : Google Scholar
5	Chen L, Song J, Cui J, Hou J, Zheng X, Li C and Liu L: microRNAs regulate adipocyte differentiation. Cell Biol Int. 37:533–546. 2013. View Article : Google Scholar : PubMed/NCBI
6	Karbiener M, Neuhold C, Oprriessnig P, Prokesch A, Bogner- Strauss JG and Scheideler M: MicroRNA-30c promotes human adipocyte differentiation and co-represses PAI-1 and ALK2. RNA Biol. 8:850–860. 2011. View Article : Google Scholar : PubMed/NCBI
7	Wang Q, Li YC, Wang J, Kong J, Qi Y, Quigg RJ and Li X: miR-17-92 cluster accelerates adipocyte differentiation by negatively regulating tumor-suppressor Rb2/p130. Proc Natl Acad Sci USA. 26:2889–2894. 2008. View Article : Google Scholar
8	Lee EK, Lee MJ, Abdelmohsen K, Kim W, Kim MM, Srikantan S, Martindale JL, Hutchison ER, Kim HH, Marasa BS, Selimyan R, Egan JM, Smith SR, Fried SK and Gorospe M: miR-130 suppresses adipogenesis by inhibiting peroxisome proliferator-activated receptor gamma expression. Mol Cell Biol. 31:626–638. 2011. View Article : Google Scholar :
9	Kim SY, Kim AY, Lee HW, Son YH, Lee GY, Lee JW, Lee YS and Kim JB: miR-27a is a negative regulator of adipocyte differentiation via suppressing PPARgamma expression. Biochem Biophys Res Commun. 392:323–328. 2010. View Article : Google Scholar : PubMed/NCBI
10	Gerin I, Bommer GT, McCoin CS, Sousa KM, Krishnan V and MacDougald OA: Roles for miRNA-378/378* in adipocyte gene expression and lipogenesis. Am J Physiol Endocrinol Metab. 299:E198–E206. 2010.PubMed/NCBI
11	Martinelli R, Nardelli C, Pilone V, Buonomo T, Liguori R, Castanò I, Buono P, Masone S, Persico G, Forestieri P, Pastore L and Sacchetti L: miR-519d overexpression is associated with human obesity. Obesity (Silver Spring). 18:2170–2176. 2010. View Article : Google Scholar
12	Kim YJ, Hwang SJ, Bae YC and Jung JS: MiR-21 regulates adipogenic differentiation through the modulation of TGF-beta signaling in mesenchymal stemcells derived from human adipose tissue. Stem Cells. 27:3093–3102. 2009.PubMed/NCBI
13	Kennell JA, Gerin I, MacDougald OA and Cadigan KM: The microRNA miR-8 is a conserved negative regulator of Wnt signaling. Proc Natl Acad Sci USA. 105:15417–1522. 2008. View Article : Google Scholar : PubMed/NCBI
14	Chen H, Wang S, Chen L, Chen Y, Wu M, Zhang Y, Yu K, Huang Z, Qin L and Mo D: MicroRNA-344 inhibits 3T3-L1 cell differentiation via targeting GSK3β of Wnt/β-catenin signaling pathway. FEBS Lett. 588:429–435. 2013. View Article : Google Scholar
15	Qin L, Chen Y, Niu Y, Chen W, Wang Q, Xiao S, Li A, Xie Y, Li J, Zhao X, He Z and Mo D: A deep investigation into the adipogenesis mechanism: profile of microRNAs regulating adipogenesis by modulating the canonical Wnt/β-catenin signaling pathway. BMC Genomics. 11:3202010. View Article : Google Scholar
16	Christodoulides C, Lagathu C, Sethi JK and Vidal-Puig A: Adipogenesis and WNT signaling. Trends Endocrinol Metab. 20:16–24. 2009. View Article : Google Scholar
17	Huang J, Zhao L, Xing L and Chen D: MicroRNA-204 regulates Runx2 protein expression and mesenchymal progenitor cell differentiation. Stem Cells. 28:357–364. 2010.
18	Yang X, Shang H, Katz A and Li X: A modified aggregate culture for chondrogenesis of human adipose-derived stem cells genetically modified with growth and differentiation factor 5. Biores Open Access. 2:258–265. 2013. View Article : Google Scholar : PubMed/NCBI
19	Hausman GJ, Dodson MV, Ajuwon K, Azain M, Barnes KM, Guan LL, Jiang Z, Poulors SP, Srainz RD, Smith S, Spurlock M, Novakofski J, Fernyhough ME and Bergen WG: Board-invited review: the biology and regulation of preadipocytes and adipocytes in meat animals. J Anim Sci. 87:1218–1246. 2009. View Article : Google Scholar
20	Chartoumpekis DV, Zaravinos A, Ziros PG, Iskrenova RP, Psyrogiannis AI, Kyriazopoulou VE and Habeos IG: Differential expression of microRNAs in adipose tissue after long-term high-fat diet-induced obesity in mice. PLoS One. 7:e348722012. View Article : Google Scholar : PubMed/NCBI
21	Fei J, Tamski H, Cook C and Santanam N: MicroRNA regulation of adipose derived stem cells in aging rats. PLoS One. 8:e592382013. View Article : Google Scholar : PubMed/NCBI
22	Zhang Y, Xie RL, Gordon J, LeBlanc K, Stein JL, Lian JB, van Wijnen AJ and Stein GS: Control of mesenchymal lineage progression by microRNAs targeting skeletal gene regulators Trps1 and Runx2. J Biol Chem. 22:21926–21935. 2012. View Article : Google Scholar
23	Liu J and Farmer SR: Regulating the balance between peroxi-some proliferator-activated receptor gamma andbeta-catenin signaling during adipogenesis. A glycogen synthase kinase 3beta phosphorylation-defective mutant of beta-catenin inhibits expression of a subset of adipogenic genes. J Biol Chem. 22:45020–45027. 2004. View Article : Google Scholar
24	Ross SE, Hemati N, Longo KA, Bennett CN, Lucas PC, Erickson RL and MacDougald OA: Inhibition of adipogenesis by Wnt signaling. Science. 11:950–953. 2000. View Article : Google Scholar
25	Lee H, Kang R, Bae S and Yoon Y: AICAR, an activator of AMPK, inhibits adipogenesis via the WNT/β-catenin pathway in 3T3-L1 adipocytes. Int J Mol Med. 28:65–71. 2011.PubMed/NCBI
26	Chan CY, Wei L, Castro-Muñozledo F and Koo WL: (−)-Epigallocatechin-3-gallate blocks 3T3-L1 adipose conversion by inhibition of cell proliferation and suppression of adipose phenotype expression. Life Sci. 21:779–785. 2011. View Article : Google Scholar
27	Lee H, Bae S and Yoon Y: The anti-adipogenic effects of (−)epigallocatechin gallate are dependent on the WNT/β-catenin pathway. J Nutr Biochem. 24:1232–1240. 2013. View Article : Google Scholar : PubMed/NCBI
28	Lee H, Bae S, Kim YS and Yoon Y: WNT/β-catenin pathway mediates the anti-adipogenic effect of platycodin D, a natural compound found in Platycodon grandiflorum. Life Sci. 12:388–394. 2011. View Article : Google Scholar
29	Lee H, Bae S and Yoon Y: Anti-adipogenic effects of 1,25-dihy-droxyvitamin D3 are mediated by the maintenance of the wingless-type MMTV integration site/β-catenin pathway. Int J Mol Med. 30:1219–1224. 2012.PubMed/NCBI
30	Lagathu C, Christodoulides C, Virtue S, Cawthorn WP, Franzin C, Kimber WA, Nora ED, Campbell M, Medina- Gomez G, Cheyette BN, Vidal-Puig AJ and Sethi JK: Dact1, a nutritionally regulated preadipocyte gene, controls adipogen-esis by coordinating the Wnt/beta-catenin signaling network. Diabetes. 58:609–619. 2009. View Article : Google Scholar :
31	Lee H, Kang R, Hahn Y, Yang Y, Kim SS, Cho SH, Chung SI and Yoon Y: Anti obesity effect of baicalin involves the modulations of proadipogenic and antiadipogenic regulators of the adipogenesis pathway. Phytother Res. 23:1615–1623. 2009. View Article : Google Scholar : PubMed/NCBI