Effects of the DUSP6 gene on the proliferation and differentiation of porcine subcutaneous preadipocytes

TL;DR

This study shows that the DUSP6 gene is important for fat cell growth and development in pigs, with higher expression in fat-type pigs.

Contribution

This study is the first to demonstrate the role of DUSP6 in porcine adipogenesis and its impact on proliferation and differentiation of preadipocytes.

Findings

DUSP6 expression is significantly higher in fat-type pigs compared to lean-type pigs in fat-related tissues.

Knockdown of DUSP6 inhibits preadipocyte proliferation and differentiation by reducing cell viability and lipid accumulation.

DUSP6 knockdown downregulates key genes and transcription factors involved in adipogenesis.

Abstract

Dual-specificity protein phosphatase 6 (DUSP6), also known as mitogen-activated protein kinase phosphatase 3 (MKP-3), was considered as a functional candidate gene for white fat accumulation in mice. However, the physiological function of the DUSP6 gene on white adipocyte adipogenesis in farm animals remains unknown. In this study, we aimed to clarify the effect of DUSP6 on porcine subcutaneous preadipocyte proliferation and differentiation.

We first make clear that the patterns of DUSP6 expression is associated with fat contents in porcine fat deposition related tissues. Porcine subcutaneous preadipocytes were isolated and induced to differentiation. Small interfering RNAs were applied to deplete DUSP6. MTT assay, CCK-8 analysis, Oil Red O staining, triglyceride determination and reverse transcription quantitative polymerase chain reaction were applied to study the regulatory role of DUSP6 during adipocyte adipogenesis in pigs.

We found that the expression levels of DUSP6 were significantly higher in backfat and longissimus dorsi tissues from fat-type pigs than in those from lean-type pigs. Consistently, the significantly induced expression of DUSP6 was also observed in differentiated adipocytes. In addition, knockdown of DUSP6 greatly inhibited preadipocytes proliferation, through the decreased cell viability and downregulated mRNA expressions of cell proliferation-associated genes, including PCNA, CDK1, CDK2. Furthermore, knockdown of DUSP6 significantly inhibited preadipocytes differentiation, as evidenced by markedly reduced lipid droplet formation, attenuated triglyceride accumulation and downregulated expression levels of adipogenic transcription masters (PPARγ, C/EBPβ, FASN and FABP4) in DUSP6 knockdown cells.

Our results demonstrate that DUSP6 is required for white adipocyte adipogenesis in pigs.

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