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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 role of Twist 1 in adipogenesis was explored using retroviral interference of Twist 1 expression. (A/B) Retroviral interference of Twist 1 expression was conducted using LV3. Western blot analysis showed that Twist 1 expression was reduced to 70-80% of the control. (C/D) Oil red O staining was used to analyze the differentiation of 3T3-L1/NC and 3T3-L1/Twist 1- cells. The results showed that interference of Twist 1 did not influence the differentiation, and there were no significant differences among the groups. (E) The expression levels of the PPARγ and ALBP proteins were increased in the Twist 1 shRNA-treated cells compared with the vector-treated control cells under hormone-induced differentiation. The infection time for both Twist 1/shRNA and LV3/NC was 48 h.
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Fig5: The role of Twist 1 in adipogenesis was explored using retroviral interference of Twist 1 expression. (A/B) Retroviral interference of Twist 1 expression was conducted using LV3. Western blot analysis showed that Twist 1 expression was reduced to 70-80% of the control. (C/D) Oil red O staining was used to analyze the differentiation of 3T3-L1/NC and 3T3-L1/Twist 1- cells. The results showed that interference of Twist 1 did not influence the differentiation, and there were no significant differences among the groups. (E) The expression levels of the PPARγ and ALBP proteins were increased in the Twist 1 shRNA-treated cells compared with the vector-treated control cells under hormone-induced differentiation. The infection time for both Twist 1/shRNA and LV3/NC was 48 h.

Mentions: Western blot analysis demonstrated that the expression of Twist 1 was 70-80% reduced in Twist 1 shRNA-treated cells compared with its expression in vector-treated control cells (Figure 5A/B). When induced to differentiate, the Twist 1 shRNA-treated cells showed no obvious differences in intracellular lipid accumulation (P > 0.05) (Figure 5C/D). The expression levels of the PPARγ and ALBP proteins were increased in the Twist 1 shRNA-treated cells compared with the vector-treated control cells under hormone-induced differentiation. (Figure 5E).Figure 5


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 role of Twist 1 in adipogenesis was explored using retroviral interference of Twist 1 expression. (A/B) Retroviral interference of Twist 1 expression was conducted using LV3. Western blot analysis showed that Twist 1 expression was reduced to 70-80% of the control. (C/D) Oil red O staining was used to analyze the differentiation of 3T3-L1/NC and 3T3-L1/Twist 1- cells. The results showed that interference of Twist 1 did not influence the differentiation, and there were no significant differences among the groups. (E) The expression levels of the PPARγ and ALBP proteins were increased in the Twist 1 shRNA-treated cells compared with the vector-treated control cells under hormone-induced differentiation. The infection time for both Twist 1/shRNA and LV3/NC was 48 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig5: The role of Twist 1 in adipogenesis was explored using retroviral interference of Twist 1 expression. (A/B) Retroviral interference of Twist 1 expression was conducted using LV3. Western blot analysis showed that Twist 1 expression was reduced to 70-80% of the control. (C/D) Oil red O staining was used to analyze the differentiation of 3T3-L1/NC and 3T3-L1/Twist 1- cells. The results showed that interference of Twist 1 did not influence the differentiation, and there were no significant differences among the groups. (E) The expression levels of the PPARγ and ALBP proteins were increased in the Twist 1 shRNA-treated cells compared with the vector-treated control cells under hormone-induced differentiation. The infection time for both Twist 1/shRNA and LV3/NC was 48 h.
Mentions: Western blot analysis demonstrated that the expression of Twist 1 was 70-80% reduced in Twist 1 shRNA-treated cells compared with its expression in vector-treated control cells (Figure 5A/B). When induced to differentiate, the Twist 1 shRNA-treated cells showed no obvious differences in intracellular lipid accumulation (P > 0.05) (Figure 5C/D). The expression levels of the PPARγ and ALBP proteins were increased in the Twist 1 shRNA-treated cells compared with the vector-treated control cells under hormone-induced differentiation. (Figure 5E).Figure 5

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