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Shp2 suppresses the adipogenic differentiation of preadipocyte 3T3-L1 cells at an early stage

View Article: PubMed Central - PubMed

ABSTRACT

Tyrosine phosphatase protein Shp2 is a potential therapeutic target for obesity. However, the mechanism of Shp2 during adipogenesis is not fully understood. The present study investigated the role of Shp2 in the terminal differentiation of preadipocytes. The results showed that Shp2 suppressed adipocyte differentiation in 3T3-L1 cells; overexpression of Shp2 reduced lipid droplet production in 3T3-L1 cells, whereas Shp2 knockdown increased lipid droplet production in 3T3-L1 cells. Furthermore, inhibition of Shp2 activity also enhanced adipocyte differentiation. Interestingly, Shp2 expression was specifically decreased early during differentiation in response to stimulation with the dexamethasone–methylisobutylxanthine–insulin (DMI) hormone cocktail. During the first 2 days of differentiation, Shp2 overexpression impaired the DMI-induced phosphorylation of signal transducer and activator of transcription 3 (STAT3) in 3T3-L1 cells and blocked the peak expression of CCAAT/enhancer-binding proteins β and δ during preadipocyte differentiation. In conclusion, Shp2 downregulated the early stages of hormone-induced differentiation of 3T3-L1 cells and inhibited the expression of the first wave of transcription factors by suppressing the DMI-induced STAT3 signaling pathway. These discoveries point to a novel role of Shp2 during adipogenesis and support the hypothesis that Shp2 could be a therapeutic target for the control of obesity.

No MeSH data available.


Shp2 suppressed the expression of C/EBPβ and δ early during 3T3-L1 adipocyte differentiation. C/EBPβ, C/EBPδ and Shp2 mRNA expression levels were measured by real-time PCR during the first 2 days of adipocyte differentiation of 3T3-L1 cells that overexpressed (a) or underexpressed (b) the Shp2 protein. The experiments were repeated at least five times, and one representative result is shown. The quantitative values are the mean±S.E.M. from all replicated experiments. The asterisks denote significance. n=3; *P<0.05; **P<0.01; ***P<0.001.
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fig6: Shp2 suppressed the expression of C/EBPβ and δ early during 3T3-L1 adipocyte differentiation. C/EBPβ, C/EBPδ and Shp2 mRNA expression levels were measured by real-time PCR during the first 2 days of adipocyte differentiation of 3T3-L1 cells that overexpressed (a) or underexpressed (b) the Shp2 protein. The experiments were repeated at least five times, and one representative result is shown. The quantitative values are the mean±S.E.M. from all replicated experiments. The asterisks denote significance. n=3; *P<0.05; **P<0.01; ***P<0.001.

Mentions: C/EBPβ and δ are part of a first wave of TFs during adipogenesis, and their expression levels peak early during preadipocyte differentiation. We found that Shp2 suppressed the expression of C/EBPβ and δ induced by DMI. The mRNA levels of these TFs were significantly decreased in 3T3-L1 cells that overexpressed Shp2 on day 1 of the differentiation process (Figure 6a, middle and right panels). Unexpectedly, C/EBPβ and δ expression recovered at day 2 (Figure 6a, middle and right panels), which may be due to the reduced expression of Shp2 protein caused by DMI (Figure 6a, left panel). Consistently, C/EBPβ expression was increased in 3T3-L1 cells with knocked down Shp2 on day 1 (Figure 6b, middle and right panels). These data suggest that Shp2 suppresses C/EBPβ and δ expression and that DMI blocks Shp2’s antagonism to increase C/EBPβ and δ mRNA levels.


Shp2 suppresses the adipogenic differentiation of preadipocyte 3T3-L1 cells at an early stage
Shp2 suppressed the expression of C/EBPβ and δ early during 3T3-L1 adipocyte differentiation. C/EBPβ, C/EBPδ and Shp2 mRNA expression levels were measured by real-time PCR during the first 2 days of adipocyte differentiation of 3T3-L1 cells that overexpressed (a) or underexpressed (b) the Shp2 protein. The experiments were repeated at least five times, and one representative result is shown. The quantitative values are the mean±S.E.M. from all replicated experiments. The asterisks denote significance. n=3; *P<0.05; **P<0.01; ***P<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4979423&req=5

fig6: Shp2 suppressed the expression of C/EBPβ and δ early during 3T3-L1 adipocyte differentiation. C/EBPβ, C/EBPδ and Shp2 mRNA expression levels were measured by real-time PCR during the first 2 days of adipocyte differentiation of 3T3-L1 cells that overexpressed (a) or underexpressed (b) the Shp2 protein. The experiments were repeated at least five times, and one representative result is shown. The quantitative values are the mean±S.E.M. from all replicated experiments. The asterisks denote significance. n=3; *P<0.05; **P<0.01; ***P<0.001.
Mentions: C/EBPβ and δ are part of a first wave of TFs during adipogenesis, and their expression levels peak early during preadipocyte differentiation. We found that Shp2 suppressed the expression of C/EBPβ and δ induced by DMI. The mRNA levels of these TFs were significantly decreased in 3T3-L1 cells that overexpressed Shp2 on day 1 of the differentiation process (Figure 6a, middle and right panels). Unexpectedly, C/EBPβ and δ expression recovered at day 2 (Figure 6a, middle and right panels), which may be due to the reduced expression of Shp2 protein caused by DMI (Figure 6a, left panel). Consistently, C/EBPβ expression was increased in 3T3-L1 cells with knocked down Shp2 on day 1 (Figure 6b, middle and right panels). These data suggest that Shp2 suppresses C/EBPβ and δ expression and that DMI blocks Shp2’s antagonism to increase C/EBPβ and δ mRNA levels.

View Article: PubMed Central - PubMed

ABSTRACT

Tyrosine phosphatase protein Shp2 is a potential therapeutic target for obesity. However, the mechanism of Shp2 during adipogenesis is not fully understood. The present study investigated the role of Shp2 in the terminal differentiation of preadipocytes. The results showed that Shp2 suppressed adipocyte differentiation in 3T3-L1 cells; overexpression of Shp2 reduced lipid droplet production in 3T3-L1 cells, whereas Shp2 knockdown increased lipid droplet production in 3T3-L1 cells. Furthermore, inhibition of Shp2 activity also enhanced adipocyte differentiation. Interestingly, Shp2 expression was specifically decreased early during differentiation in response to stimulation with the dexamethasone&ndash;methylisobutylxanthine&ndash;insulin (DMI) hormone cocktail. During the first 2 days of differentiation, Shp2 overexpression impaired the DMI-induced phosphorylation of signal transducer and activator of transcription 3 (STAT3) in 3T3-L1 cells and blocked the peak expression of CCAAT/enhancer-binding proteins &beta; and &delta; during preadipocyte differentiation. In conclusion, Shp2 downregulated the early stages of hormone-induced differentiation of 3T3-L1 cells and inhibited the expression of the first wave of transcription factors by suppressing the DMI-induced STAT3 signaling pathway. These discoveries point to a novel role of Shp2 during adipogenesis and support the hypothesis that Shp2 could be a therapeutic target for the control of obesity.

No MeSH data available.