UHRF1 Promotes Proliferation of Human Adipose-Derived Stem Cells and Suppresses Adipogenesis via Inhibiting Peroxisome Proliferator-Activated Receptor γ.

Ke Chen,Zhaohui Mo,Yufang Luo,Zi Guo,Jingjing Yuan

doi:10.1155/2019/9456847

Abstract

Once the adipose tissue is enlarged for the purpose of saving excess energy intake, obesity may be observed. Ubiquitin-like with PHD and RING Finger domains 1 (UHRF1) is helpful in repairing damaged DNA as it increases the resistance of cancer cells against cytocidal drugs. Peroxisome proliferator-activated receptor γ (PPARγ), an important nucleus transcription factor participating in adipogenesis, has been extensively reported. To date, no study has indicated whether UHRF1 can regulate proliferation and differentiation of human adipose-derived stem cells (hADSCs). Hence, this study aimed to utilize overexpression or downregulation of UHRF1 to explore the possible mechanism of proliferation and differentiation of hADSCs. We here used lentivirus, containing UHRF1 (LV-UHRF1) and siRNA-UHRF1 to transfect hADSCs, on which Cell Counting Kit-8 (CCK-8), cell growth curve, colony formation assay, and EdU proliferation assay were applied to evaluate proliferation of hADSCs, cells cycle was investigated by flow cytometry, and adipogenesis was detected by Oil Red O staining and Western blotting. Our results showed that UHRF1 can promote proliferation of hADSCs after overexpression of UHRF1, while proliferation of hADSCs was reduced through downregulation of UHRF1, and UHRF1 can control proliferation of hADSCs through transition from G1-phase to S-phase; besides, we found that UHRF1 negatively regulates adipogenesis of hADSCs via PPARγ. In summary, the results may provide a new insight regarding the role of UHRF1 on regulating proliferation and differentiation of hADSCs.

Highlights

Obesity may lead to a series of serious metabolism diseases, such as hypertension, diabetes, cardiovascular disease, and dyslipidemia [1]
We first detected the identification and characterization of human adipose-derived stem cells (hADSCs), and our results showed that the typical surface marker of mesenchymal stem cells (MSCs) was expressed in hADSCs
We analyzed the expression of peroxisome proliferator-activated receptor γ (PPARγ) and C/EBPα mRNA after lentivirus negative control (LV-NC), LV-UHRF1, siRNAUHRF1, and Small Interfering RNA (siRNA)-NC were transfected into hADSCs for 3 days, respectively, and we found that overexpression of UHRF1 could inhibit expression of PPARγ, whereas downregulation of UHRF1 could promote expression of PPARγ (∗P < 0.05,△P < 0.01 compared with LV-NC group; Figures 4(f) and 4(g))

Summary

Introduction

Obesity may lead to a series of serious metabolism diseases, such as hypertension, diabetes, cardiovascular disease, and dyslipidemia [1]. UHRF1 is able to regulate DNA-methylation via different DNA-binding proteins, BioMed Research International such as histone H3 lysine 9 (H3K9), histone deacetylase 1 (HDAC1), DNA methyltransferase 1 (DNMT1), proliferating cell nuclear antigen (PCNA), and euchromatic histone-lysine N-methyltransferase 2 (EHMT2)[12, 13]. An another important function of UHRF1 is to promote cell proliferation, which has been extensively reported [14,15,16]

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Results

Conclusion