Obesity‑associated microRNA‑26b regulates the proliferation of human preadipocytes via arrest of the G1/S transition

Xu,Guangfeng; Ji,Chenbo; Song,Guixian; Shi,Chunmei; Shen,Yahui; Chen,Ling; Yang,Lei; Zhao,Yaping; Guo,Xirong

doi:10.3892/mmr.2015.3858

Obesity‑associated microRNA‑26b regulates the proliferation of human preadipocytes via arrest of the G1/S transition

Authors:
- Guangfeng Xu
- Chenbo Ji
- Guixian Song
- Chunmei Shi
- Yahui Shen
- Ling Chen
- Lei Yang
- Yaping Zhao
- Xirong Guo
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Affiliations: Department of Laboratory Medicine, The 82nd Hospital of The People's Liberation Army, Huaian, Jiangsu 223001, P.R. China, Department of Child Health Care, Affiliated Nanjing Maternity and Child Health Care Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China, Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China, Institute of Pediatrics, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China
Published online on: May 27, 2015 https://doi.org/10.3892/mmr.2015.3858
Pages: 3648-3654

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Abstract

MicroRNAs (miRNAs) are short, 20‑24 nucleotide non‑coding RNAs, which are involved in multiple biological processes, including obesity. Our previous investigation revealed that miRNA (miR)‑26b is differentially expressed in preadipocytes and mature adipocytes in humans. However, its role in the proliferation of human preadipocytes remains to be fully elucidated. In the present study, intracellular lipid accumulation was assessed using oil red O staining and the trigycerlide (TG) content was quantified using a TG assay kit, adipogenesis associated genes and cyclin D2 were analyzed using western blotting, and the effects of miR‑26b on the proliferation of preadipocytes was investigated using Cell Counting Kit‑8 assays and cell cycle analysis. Human preadipocytes overexpressing miR‑26b exhibited increased TG content in the adipocytes. During differentiation, the protein expression levels of adipogenesis‑associated marker genes, including peroxisome proliferator‑activated receptor γ, CCAAT/enhancer‑binding protein α, fatty acid‑binding protein and hormone‑sensitive lipase were upregulated in cells overexpressing miR‑26b, compared with the negative control cells. In addition, growth of human preadipocytes overexpressing miR‑26b occurred a slower rate and more remained in the G1 phase, compared with the negative control cells. In addition, miR‑26b downregulated the protein expression of cyclin D2. These results demonstrated that miR‑26b promoted differentiation and, at least party by targeting cyclin D2, attenuated cell proliferation via arresting the G1/S transition.

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1	Haslam DW and James WP: Obesity. Lancet. 366:1197–1209. 2005. View Article : Google Scholar : PubMed/NCBI
2	Basham P and Luik J: Is the obesity epidemic exaggerated? Yes. BMJ. 336:2442008. View Article : Google Scholar : PubMed/NCBI
3	Avram MM, Avram AS and James WD: Subcutaneous fat in normal and diseased states 3. Adipogenesis: from stem cell to fat cell. J Am Acad Dermatol. 56:472–492. 2007. View Article : Google Scholar : PubMed/NCBI
4	Hausman DB, DiGirolamo M, Bartness TJ, Hausman GJ and Martin RJ: The biology of white adipocyte proliferation. Obes Rev. 2:239–254. 2001. View Article : Google Scholar
5	Bushati N and Cohen SM: microRNA functions. Annu Rev Cell Dev Biol. 23:175–205. 2007. View Article : Google Scholar : PubMed/NCBI
6	Eulalio A, Huntzinger E and Izaurralde E: Getting to the root of miRNA-mediated gene silencing. Cell. 132:9–14. 2008. View Article : Google Scholar : PubMed/NCBI
7	Kim YJ, Hwang SH, Cho HH, Shin KK, Bae YC and Jung JS: MicroRNA 21 regulates the proliferation of human adipose tissue-derived mesenchymal stem cells and high-fat diet-induced obesity alters microRNA 21 expression in white adipose tissues. J Cell Physiol. 227:183–193. 2012. View Article : Google Scholar
8	Zhang JF, Fu WM, He ML, et al: MiR-637 maintains the balance between adipocytes and osteoblasts by directly targeting Osterix. Mol Biol Cell. 22:3955–3961. 2011. View Article : Google Scholar : PubMed/NCBI
9	Esau C, Kang X, Peralta E, et al: MicroRNA-143 regulates adipocyte differentiation. J Biol Chem. 279:52361–52365. 2004. View Article : Google Scholar : PubMed/NCBI
10	Karbiener M, Neuhold C, Opriessnig 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
11	Lee EK, Lee MJ, Abdelmohsen K, et al: miR-130 suppresses adipogenesis by inhibiting peroxisome proliferator-activated receptor gamma expression. Mol Cell Biol. 31:626–638. 2011. View Article : Google Scholar :
12	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using realtime quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
13	Dill H, Linder B, Fehr A and Fischer U: Intronic miR-26b controls neuronal differentiation by repressing its host transcript, ctdsp2. Genes Dev. 26:25–30. 2012. View Article : Google Scholar : PubMed/NCBI
14	Song G, Xu G, Ji C, et al: The role of microRNA-26b in human adipocyte differentiation and proliferation. Gene. 533:481–487. 2014. View Article : Google Scholar
15	Xu G, Ji C, Shi C, et al: Modulation of hsa-miR-26b levels following adipokine stimulation. Mol Biol Rep. 40:3577–3582. 2013. View Article : Google Scholar : PubMed/NCBI
16	Xu G, Shi C, Ji C, et al: Expression of microRNA-26b, an obesity-related microRNA, is regulated by free fatty acids, glucose, dexamethasone and growth hormone in human adipocytes. Mol Med Rep. 10:223–228. 2014.PubMed/NCBI
17	Klöting N, Berthold S, Kovacs P, et al: MicroRNA expression in human omental and subcutaneous adipose tissue. PLoS One. 4:e46992009. View Article : Google Scholar : PubMed/NCBI
18	Kajimoto K, Naraba H and Iwai N: MicroRNA and 3T3-L1 pre-adipocyte differentiation. Rna. 12:1626–1632. 2006. View Article : Google Scholar : PubMed/NCBI
19	Sun F, Wang J, Pan Q, et al: Characterization of function and regulation of miR-24-1 and miR-31. Biochem Biophys Res Commun. 380:660–665. 2009. View Article : Google Scholar : PubMed/NCBI
20	Jones JR, Barrick C, Kim KA, et al: Deletion of PPARgamma in adipose tissues of mice protects against high fat diet-induced obesity and insulin resistance. Proc Natl Acad Sci USA. 102:6207–6212. 2005. View Article : Google Scholar : PubMed/NCBI
21	Zuo Y, Qiang L and Farmer SR: Activation of CCAAT/enhancer-binding protein (C/EBP) alpha expression by C/EBP beta during adipogenesis requires a peroxisome proliferator-activated receptor-gamma-associated repression of HDAC1 at the C/ebp alpha gene promoter. J Biol Chem. 281:7960–7967. 2006. View Article : Google Scholar : PubMed/NCBI
22	Kralisch S and Fasshauer M: Adipocyte fatty acid binding protein: a novel adipokine involved in the pathogenesis of metabolic and vascular disease? Diabetologia. 56:10–21. 2013. View Article : Google Scholar
23	Della-Fera MA, Qian H and Baile CA: Adipocyte apoptosis in the regulation of body fat mass by leptin. Diabetes Obes Metab. 3:299–310. 2001. View Article : Google Scholar : PubMed/NCBI