Differential microRNA expression profile comparison between epidermal stem cells and differentiated keratinocytes

Song,Zhifang; Liu,Dewu; Peng,Yan; Li,Jin; Zhang,Zhiwei; Ning,Pu

doi:10.3892/mmr.2014.2886

Differential microRNA expression profile comparison between epidermal stem cells and differentiated keratinocytes

Authors:
- Zhifang Song
- Dewu Liu
- Yan Peng
- Jin Li
- Zhiwei Zhang
- Pu Ning
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Affiliations: Department of Cardiovascular Medicine, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Burn Center, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
Published online on: November 6, 2014 https://doi.org/10.3892/mmr.2014.2886
Pages: 2285-2291

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Abstract

The aim of the current study was to analyze the differential microRNA (miRNA) expression profiles of human epidermal stem cells (ESCs) and differentiated keratinocytes. Enzyme digestion was used in combination with rapid adhesion to collagen IV to isolate primary human ESCs and differentiated keratinocytes, from which total RNA was extracted. Fluorescence labeling, microarray hybridization and differential expression analyses were performed. Reverse transcription quantitative polymerase chain reaction (RT‑qPCR) was performed to validate the reliability of the microarray results and predict the target genes of the differentially expressed miRNAs. A total of 25 miRNAs, including hsa‑miR‑197‑5p, hsa‑miR‑125b‑5p and hsa‑miR‑376a‑3p, were upregulated, whereas 166 miRNAs, including hsa‑miR‑29b‑3p, hsa‑miR‑203 and hsa‑miR‑34a‑3p, were downregulated in the human ESCs compared with the expression in differentiated keratinocytes. RT‑qPCR results confirmed the upregulation of hsa‑miR‑197‑5p and the downregulation of hsa‑miR‑29b‑3p, which were consistent with the microarray results. miRNA target prediction indicated that the miRNA expression levels correlated with cell proliferation, differentiation, apoptosis and senescence. Expression levels of miRNAs significantly differed between human ESCs and differentiated keratinocytes. This finding may be attributed to their biological characteristics, such as proliferative behavior and differentiation abilities.

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1	Sandilands A, Sutherland C, Irvine AD and McLean WH: Filaggrin in the frontline: role in skin barrier function and disease. J Cell Sci. 122:1285–1294. 2009. View Article : Google Scholar : PubMed/NCBI
2	Sotiropoulou PA and Blanpain C: Development and homeostasis of the skin epidermis. Cold Spring Harb Perspect Biol. 4:a0083832012. View Article : Google Scholar : PubMed/NCBI
3	Cangkrama M, Ting SB and Darido C: Stem cells behind the barrier. Int J Mol Sci. 14:13670–13686. 2013. View Article : Google Scholar : PubMed/NCBI
4	Moreno-Moya JM, Vilella F and Simón C: MicroRNA: key gene expression regulators. Fertil Steril. 101:1516–1523. 2014. View Article : Google Scholar
5	Cai Y, Yu X, Hu S and Yu J: A brief review on the mechanisms of miRNA regulation. Genomics Proteomics Bioinformatics. 7:147–154. 2009. View Article : Google Scholar
6	Andl T, Murchison EP, Liu F, et al: The miRNA-processing enzyme dicer is essential for the morphogenesis and maintenance of hair follicles. Curr Biol. 16:1041–1049. 2006. View Article : Google Scholar : PubMed/NCBI
7	Yi R, Pasolli HA, Landthaler M, et al: DGCR8-dependent microRNA biogenesis is essential for skin development. Proc Natl Acad Sci USA. 106:498–502. 2009. View Article : Google Scholar :
8	Kim DS, Cho HJ, Choi HR, Kwon SB and Park KC: Isolation of human epidermal stem cells by adherence and the reconstruction of skin equivalents. Cell Mol Life Sci. 61:2774–2781. 2004. View Article : Google Scholar : PubMed/NCBI
9	Schmittgen TD and Livak KJ: Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 3:1101–1108. 2008. View Article : Google Scholar : PubMed/NCBI
10	Lewis BP, Burge CB and Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 120:15–20. 2005. View Article : Google Scholar : PubMed/NCBI
11	Yi R and Fuchs E: MicroRNA-mediated control in the skin. Cell Death Differ. 17:229–235. 2010. View Article : Google Scholar :
12	Ghadially R: 25 years of epidermal stem cell research. J Invest Dermatol. 132:797–810. 2012. View Article : Google Scholar
13	Hildebrand J, Rütze M, Walz N, et al: A comprehensive analysis of microRNA expression during human keratinocyte differentiation in vitro and in vivo. J Invest Dermatol. 131:20–29. 2011. View Article : Google Scholar
14	Hildebrand J, Grundhoff A, Gallinat S, et al: MicroRNA profiling during human keratinocyte differentiation using a quantitative real-time PCR method. Methods Mol Biol. 961:193–200. 2013. View Article : Google Scholar : PubMed/NCBI
15	Yi R, O’Carroll D, Pasolli HA, et al: Morphogenesis in skin is governed by discrete sets of differentially expressed microRNAs. Nat Genet. 38:356–362. 2006. View Article : Google Scholar : PubMed/NCBI
16	Sonkoly E, Wei T, Janson PC, et al: MicroRNAs: novel regulators involved in the pathogenesis of psoriasis? PLoS One. 2:e6102007. View Article : Google Scholar : PubMed/NCBI
17	Yi R, Poy MN, Stoffel M and Fuchs E: A skin microRNA promotes differentiation by repressing ‘stemness’. Nature. 452:225–229. 2008. View Article : Google Scholar : PubMed/NCBI
18	Johnson CD, Esquela-Kerscher A, Stefani G, et al: The let-7 microRNA represses cell proliferation pathways in human cells. Cancer Res. 67:7713–7722. 2007. View Article : Google Scholar : PubMed/NCBI
19	Peng H, Kaplan N, Hamanaka RB, et al: microRNA-31/factor-inhibiting hypoxia-inducible factor 1 nexus regulates keratinocyte differentiation. Proc Natl Sci USA. 109:14030–14034. 2012. View Article : Google Scholar
20	Mardaryev AN, Ahmed MI, Vlahov NV, et al: Micro-RNA-31 controls hair cycle-associated changes in gene expression programs of the skin and hair follicle. FASEB J. 24:3869–3881. 2010. View Article : Google Scholar : PubMed/NCBI
21	Biswas S, Roy S, Banerjee J, et al: Hypoxia inducible microRNA 210 attenuates keratinocyte proliferation and impairs closure in a murine model of ischemic wounds. Proc Natl Acad Sci USA. 107:6976–6981. 2010. View Article : Google Scholar : PubMed/NCBI
22	Bertero T, Gastaldi C, Bourget-Ponzio I, et al: miR-483–3p controls proliferation in wounded epithelial cells. FASEB J. 25:3092–3105. 2011. View Article : Google Scholar : PubMed/NCBI
23	Shin KH, Pucar A, Kim RH, et al: Identification of senescence-inducing microRNAs in normal human keratinocytes. Int J Oncol. 39:1205–1211. 2011.PubMed/NCBI
24	Lena AM, Mancini M, Rivetti di Val Cervo P, et al: MicroRNA-191 triggers keratinocytes senescence by SATB1 and CDK6 downregulation. Biochem Biophys Res Commun. 423:509–514. 2012. View Article : Google Scholar : PubMed/NCBI
25	Zhang L, Stokes N, Polak L and Fuchs E: Specific microRNAs are preferentially expressed by skin stem cells to balance self-renewal and early lineage commitment. Cell Stem Cell. 8:294–308. 2011. View Article : Google Scholar : PubMed/NCBI
26	Giglio S, Cirombella R, Amodeo R, et al: MicroRNA miR-24 promotes cell proliferation by targeting the CDKs inhibitors p27Kip1 and p16INK4a. J Cell Physiol. 228:2015–2023. 2013. View Article : Google Scholar : PubMed/NCBI
27	Yang X, Wang J, Guo SL, et al: miR-21 promotes keratinocyte migration and re-epithelialization during wound healing. Int J Biol Sci. 7:685–690. 2011. View Article : Google Scholar : PubMed/NCBI
28	Yu J, Peng H, Ruan Q, et al: MicroRNA-205 promotes keratinocyte migration via the lipid phosphatase SHIP2. FASEB J. 24:3950–3959. 2010. View Article : Google Scholar : PubMed/NCBI