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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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3T3-L1 preadipocytes were induced to differentiate in vitro, and the dynamic expression of Twist 1 was detected. (A) Morphological observations and oil red O staining of 3T3-L1 preadipocytes and adipocytes. (B) Quantitative analysis of intracellular oil red O staining using spectrophotometry. (C) PPARγ and ALBP protein expression levels at different stages of 3T3-L1 preadipocyte differentiation. (D/F) The dynamic transcription of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells). (E/G) The dynamic expression of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells).
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Fig4: 3T3-L1 preadipocytes were induced to differentiate in vitro, and the dynamic expression of Twist 1 was detected. (A) Morphological observations and oil red O staining of 3T3-L1 preadipocytes and adipocytes. (B) Quantitative analysis of intracellular oil red O staining using spectrophotometry. (C) PPARγ and ALBP protein expression levels at different stages of 3T3-L1 preadipocyte differentiation. (D/F) The dynamic transcription of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells). (E/G) The dynamic expression of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells).

Mentions: Morphological observations revealed that the lipid droplets were distributed as a ring in the cytoplasm. Oil red O staining showed clear red dye in the adipocytes; however, this dye was not observed in the preadipocytes (Figure 4A). Quantitative analysis of the intracellular oil red O by spectrophotometry demonstrated the accumulation of lipids (Figure 4B). The levels of PPARγ and ALBP were significantly increased beginning at day 2 post-differentiation induction, and the levels peaked on day 12 (Figure 4C) (P < 0.05).Figure 4


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)

3T3-L1 preadipocytes were induced to differentiate in vitro, and the dynamic expression of Twist 1 was detected. (A) Morphological observations and oil red O staining of 3T3-L1 preadipocytes and adipocytes. (B) Quantitative analysis of intracellular oil red O staining using spectrophotometry. (C) PPARγ and ALBP protein expression levels at different stages of 3T3-L1 preadipocyte differentiation. (D/F) The dynamic transcription of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells). (E/G) The dynamic expression of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: 3T3-L1 preadipocytes were induced to differentiate in vitro, and the dynamic expression of Twist 1 was detected. (A) Morphological observations and oil red O staining of 3T3-L1 preadipocytes and adipocytes. (B) Quantitative analysis of intracellular oil red O staining using spectrophotometry. (C) PPARγ and ALBP protein expression levels at different stages of 3T3-L1 preadipocyte differentiation. (D/F) The dynamic transcription of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells). (E/G) The dynamic expression of Twist 1 was detected at different stages of 3T3-L1 preadipocyte differentiation (*P < 0.05 vs. control, day 0 cells).
Mentions: Morphological observations revealed that the lipid droplets were distributed as a ring in the cytoplasm. Oil red O staining showed clear red dye in the adipocytes; however, this dye was not observed in the preadipocytes (Figure 4A). Quantitative analysis of the intracellular oil red O by spectrophotometry demonstrated the accumulation of lipids (Figure 4B). The levels of PPARγ and ALBP were significantly increased beginning at day 2 post-differentiation induction, and the levels peaked on day 12 (Figure 4C) (P < 0.05).Figure 4

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