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Twist 1 regulates the expression of PPARγ during hormone-induced 3T3-L1 preadipocyte differentiation: a possible role in obesity and associated diseases.

Ma W, Lu S, Sun T, Wang X, Ma Y, Zhang X, Zhao R, Wang Y - Lipids Health Dis (2014)

Bottom Line: Twist 1 mRNA and protein levels were reduced in diet-induced obese mice and rats and in obese humans.Retroviral interference of Twist 1 expression did not significantly impair lipid formation; however, retroviral interference induced PPARγ mRNA and protein expression on day 4 of differentiation induction.Adipokine array analyses revealed increased secretion of CXCR4 (19.55-fold), VEGFR1 (92.13-fold), L-21 R (63.55-fold), and IL-12 R beta 1 (59.66-fold) and decreased secretion of VEGFR3 (0.01-fold), TSLP R (0.071-fold), MIP-1 gamma (0.069-fold), TNF RI/TNFRSF1A (0.09-fold), and MFG-E8 (0.06-fold).

View Article: PubMed Central - PubMed

Affiliation: Medical Research & Laboratory Diagnostic Center, Jinan Center Hospital Affiliated to Shandong University, Jinan, Shandong 250013, P,R, China. mwsqianyi@163.com.

ABSTRACT

Background: Twist 1 is highly expressed in adipose tissue and has been associated with obesity and related disorders. However, the molecular function of Twist 1 in adipose tissue is unclear. Twist 1 has been implicated in cell lineage determination and differentiation. Therefore, we investigated both the role of Twist 1 in adipocyte precursor mobilization and the relationship of Twist 1 with other molecular determinants of adipocyte differentiation.

Methods: We examined Twist 1 mRNA and protein expression in subcutaneous adipose tissues from diet-induced obese C57/BL6 mice and Wistar rats and in obese patients undergoing liposuction or adipose transplant surgeries. Twist 1 expression was measured on days 0, 2, 4, 8, and 12 of 3T3-L1 differentiation in vitro. The role of Twist 1 in adipogenesis was explored using retroviral interference of Twist 1 expression. Adipokine secretion was evaluated using a RayBio® Biotin Label-based Adipokine Array.

Results: Twist 1 mRNA and protein levels were reduced in diet-induced obese mice and rats and in obese humans. Twist 1 was upregulated during 3T3-L1 preadipocyte differentiation in vitro, beginning from the fourth day of differentiation induction. Retroviral interference of Twist 1 expression did not significantly impair lipid formation; however, retroviral interference induced PPARγ mRNA and protein expression on day 4 of differentiation induction. Adipokine array analyses revealed increased secretion of CXCR4 (19.55-fold), VEGFR1 (92.13-fold), L-21 R (63.55-fold), and IL-12 R beta 1 (59.66-fold) and decreased secretion of VEGFR3 (0.01-fold), TSLP R (0.071-fold), MIP-1 gamma (0.069-fold), TNF RI/TNFRSF1A (0.09-fold), and MFG-E8 (0.06-fold).

Conclusions: Twist 1 is a regulator of adipocyte gene expression although it is not likely to regulate differentiation. We identified PPARγ as a potential target of Twist 1 and found variation in the secretion of multiple adipokines, which might indicate a prospective mechanism linking Twist 1 expression with obesity or associated diseases.

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The secretion of various adipokines by cultured 3T3-L1 adipocytes with or without Twist 1 interference was evaluated using a Mouse Antibody Array. (A) Data from the array analyses with filter 1, which was treated with medium from the adipocytes infected with LV3/NC for 48 h (Left), and filter 2, which was treated with medium from the adipocytes infected with Twist 1/shRNA for 48 h (Right). (B) The values obtained from the scans were adjusted based on the intensity of the control spots on the filter corners, and the elevated levels of specific adipokines are shown. (C) Adipokines that were decreased by more than five fold in the medium of Twist 1-knockdown cells are listed.
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Fig7: The secretion of various adipokines by cultured 3T3-L1 adipocytes with or without Twist 1 interference was evaluated using a Mouse Antibody Array. (A) Data from the array analyses with filter 1, which was treated with medium from the adipocytes infected with LV3/NC for 48 h (Left), and filter 2, which was treated with medium from the adipocytes infected with Twist 1/shRNA for 48 h (Right). (B) The values obtained from the scans were adjusted based on the intensity of the control spots on the filter corners, and the elevated levels of specific adipokines are shown. (C) Adipokines that were decreased by more than five fold in the medium of Twist 1-knockdown cells are listed.

Mentions: As shown in Figure 7, the levels of twenty adipokines were increased by more than two-fold in the media of the Twist 1 shRNA-treated cells, including vascular endothelial growth factor receptor 1 (VEGFR1, 92.13-fold), interleukin 21 receptor (IL-21 R, 63.55-fold), interleukin 12 receptor beta 1 (IL-12 R beta 1, 59.66-fold), Cys-X-Cys receptor 4 (CXCR4, 19.55-fold), fibroblast growth factor receptor 3 (FGF R3, 29.79-fold), and growth hormone receptor (GHR, 5.94-fold) (Figure 7B).The levels of a total of fifty-seven adipokines were decreased. There was a greater than 5-fold change observed in the adipokines shown in Figure 7C, including VEGFR3 (0.01-fold), thymic stromal lymphopoietin receptor (TSLP R, 0.071-fold), macrophage inflammatory protein 1 gamma (MIP-1 gamma, 0.069-fold), TNF RI/TNFRSF1A (0.09-fold), milk fat globule EGF factor 8 (MFG-E8, 0.06-fold), IGFBP-rp1/IGFBP-7 (0.04-fold), and CD27 ligand/TNFSF7 (0.09-fold).Figure 7


Twist 1 regulates the expression of PPARγ during hormone-induced 3T3-L1 preadipocyte differentiation: a possible role in obesity and associated diseases.

Ma W, Lu S, Sun T, Wang X, Ma Y, Zhang X, Zhao R, Wang Y - Lipids Health Dis (2014)

The secretion of various adipokines by cultured 3T3-L1 adipocytes with or without Twist 1 interference was evaluated using a Mouse Antibody Array. (A) Data from the array analyses with filter 1, which was treated with medium from the adipocytes infected with LV3/NC for 48 h (Left), and filter 2, which was treated with medium from the adipocytes infected with Twist 1/shRNA for 48 h (Right). (B) The values obtained from the scans were adjusted based on the intensity of the control spots on the filter corners, and the elevated levels of specific adipokines are shown. (C) Adipokines that were decreased by more than five fold in the medium of Twist 1-knockdown cells are listed.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150960&req=5

Fig7: The secretion of various adipokines by cultured 3T3-L1 adipocytes with or without Twist 1 interference was evaluated using a Mouse Antibody Array. (A) Data from the array analyses with filter 1, which was treated with medium from the adipocytes infected with LV3/NC for 48 h (Left), and filter 2, which was treated with medium from the adipocytes infected with Twist 1/shRNA for 48 h (Right). (B) The values obtained from the scans were adjusted based on the intensity of the control spots on the filter corners, and the elevated levels of specific adipokines are shown. (C) Adipokines that were decreased by more than five fold in the medium of Twist 1-knockdown cells are listed.
Mentions: As shown in Figure 7, the levels of twenty adipokines were increased by more than two-fold in the media of the Twist 1 shRNA-treated cells, including vascular endothelial growth factor receptor 1 (VEGFR1, 92.13-fold), interleukin 21 receptor (IL-21 R, 63.55-fold), interleukin 12 receptor beta 1 (IL-12 R beta 1, 59.66-fold), Cys-X-Cys receptor 4 (CXCR4, 19.55-fold), fibroblast growth factor receptor 3 (FGF R3, 29.79-fold), and growth hormone receptor (GHR, 5.94-fold) (Figure 7B).The levels of a total of fifty-seven adipokines were decreased. There was a greater than 5-fold change observed in the adipokines shown in Figure 7C, including VEGFR3 (0.01-fold), thymic stromal lymphopoietin receptor (TSLP R, 0.071-fold), macrophage inflammatory protein 1 gamma (MIP-1 gamma, 0.069-fold), TNF RI/TNFRSF1A (0.09-fold), milk fat globule EGF factor 8 (MFG-E8, 0.06-fold), IGFBP-rp1/IGFBP-7 (0.04-fold), and CD27 ligand/TNFSF7 (0.09-fold).Figure 7

Bottom Line: Twist 1 mRNA and protein levels were reduced in diet-induced obese mice and rats and in obese humans.Retroviral interference of Twist 1 expression did not significantly impair lipid formation; however, retroviral interference induced PPARγ mRNA and protein expression on day 4 of differentiation induction.Adipokine array analyses revealed increased secretion of CXCR4 (19.55-fold), VEGFR1 (92.13-fold), L-21 R (63.55-fold), and IL-12 R beta 1 (59.66-fold) and decreased secretion of VEGFR3 (0.01-fold), TSLP R (0.071-fold), MIP-1 gamma (0.069-fold), TNF RI/TNFRSF1A (0.09-fold), and MFG-E8 (0.06-fold).

View Article: PubMed Central - PubMed

Affiliation: Medical Research & Laboratory Diagnostic Center, Jinan Center Hospital Affiliated to Shandong University, Jinan, Shandong 250013, P,R, China. mwsqianyi@163.com.

ABSTRACT

Background: Twist 1 is highly expressed in adipose tissue and has been associated with obesity and related disorders. However, the molecular function of Twist 1 in adipose tissue is unclear. Twist 1 has been implicated in cell lineage determination and differentiation. Therefore, we investigated both the role of Twist 1 in adipocyte precursor mobilization and the relationship of Twist 1 with other molecular determinants of adipocyte differentiation.

Methods: We examined Twist 1 mRNA and protein expression in subcutaneous adipose tissues from diet-induced obese C57/BL6 mice and Wistar rats and in obese patients undergoing liposuction or adipose transplant surgeries. Twist 1 expression was measured on days 0, 2, 4, 8, and 12 of 3T3-L1 differentiation in vitro. The role of Twist 1 in adipogenesis was explored using retroviral interference of Twist 1 expression. Adipokine secretion was evaluated using a RayBio® Biotin Label-based Adipokine Array.

Results: Twist 1 mRNA and protein levels were reduced in diet-induced obese mice and rats and in obese humans. Twist 1 was upregulated during 3T3-L1 preadipocyte differentiation in vitro, beginning from the fourth day of differentiation induction. Retroviral interference of Twist 1 expression did not significantly impair lipid formation; however, retroviral interference induced PPARγ mRNA and protein expression on day 4 of differentiation induction. Adipokine array analyses revealed increased secretion of CXCR4 (19.55-fold), VEGFR1 (92.13-fold), L-21 R (63.55-fold), and IL-12 R beta 1 (59.66-fold) and decreased secretion of VEGFR3 (0.01-fold), TSLP R (0.071-fold), MIP-1 gamma (0.069-fold), TNF RI/TNFRSF1A (0.09-fold), and MFG-E8 (0.06-fold).

Conclusions: Twist 1 is a regulator of adipocyte gene expression although it is not likely to regulate differentiation. We identified PPARγ as a potential target of Twist 1 and found variation in the secretion of multiple adipokines, which might indicate a prospective mechanism linking Twist 1 expression with obesity or associated diseases.

Show MeSH
Related in: MedlinePlus