Limits...
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.

Show MeSH

Related in: MedlinePlus

Retroviral interference of Twist 1 expression enhanced the expression of PPARγ on day 4 of hormone-induced differentiation. (A) Real-time PCR analysis revealed that PPARγ mRNA was upregulated on day 4 of hormone-induced differentiation after retroviral interference of Twist 1 expression compared with its expression level in 3T3-L1/NC cells (*P < 0.05 vs. day 0). (B/C) PPARγ protein expression on day 4 of hormone-induced differentiation was significantly enhanced when Twist 1 expression was downregulated; there were no obvious differences at the other time points (*P < 0.05 vs. 3T3-L1/NC cells at day 4). 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
getmorefigures.php?uid=PMC4150960&req=5

Fig6: Retroviral interference of Twist 1 expression enhanced the expression of PPARγ on day 4 of hormone-induced differentiation. (A) Real-time PCR analysis revealed that PPARγ mRNA was upregulated on day 4 of hormone-induced differentiation after retroviral interference of Twist 1 expression compared with its expression level in 3T3-L1/NC cells (*P < 0.05 vs. day 0). (B/C) PPARγ protein expression on day 4 of hormone-induced differentiation was significantly enhanced when Twist 1 expression was downregulated; there were no obvious differences at the other time points (*P < 0.05 vs. 3T3-L1/NC cells at day 4). The infection time for both Twist 1/shRNA and LV3/NC was 48 h.

Mentions: The mRNA level of PPARγ was assayed by real time PCR, and the results are expressed as the fold change compared to the control. The data were analyzed using the 2-ΔΔ Ct method. The results confirmed the upregulation of PPARγ on day 4 of differentiation induction (P < 0.05); however, there were no obvious differences at the other time points (P > 0.05) (Figure 6A). A western blot assay confirmed the increased expression of PPARγ on day 4 of differentiation induction (P < 0.05), although there was no obvious difference at the other time points (P > 0.05) (Figure 6B/C). These results were consistent with the results of the real-time PCR.Figure 6


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)

Retroviral interference of Twist 1 expression enhanced the expression of PPARγ on day 4 of hormone-induced differentiation. (A) Real-time PCR analysis revealed that PPARγ mRNA was upregulated on day 4 of hormone-induced differentiation after retroviral interference of Twist 1 expression compared with its expression level in 3T3-L1/NC cells (*P < 0.05 vs. day 0). (B/C) PPARγ protein expression on day 4 of hormone-induced differentiation was significantly enhanced when Twist 1 expression was downregulated; there were no obvious differences at the other time points (*P < 0.05 vs. 3T3-L1/NC cells at day 4). 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

Fig6: Retroviral interference of Twist 1 expression enhanced the expression of PPARγ on day 4 of hormone-induced differentiation. (A) Real-time PCR analysis revealed that PPARγ mRNA was upregulated on day 4 of hormone-induced differentiation after retroviral interference of Twist 1 expression compared with its expression level in 3T3-L1/NC cells (*P < 0.05 vs. day 0). (B/C) PPARγ protein expression on day 4 of hormone-induced differentiation was significantly enhanced when Twist 1 expression was downregulated; there were no obvious differences at the other time points (*P < 0.05 vs. 3T3-L1/NC cells at day 4). The infection time for both Twist 1/shRNA and LV3/NC was 48 h.
Mentions: The mRNA level of PPARγ was assayed by real time PCR, and the results are expressed as the fold change compared to the control. The data were analyzed using the 2-ΔΔ Ct method. The results confirmed the upregulation of PPARγ on day 4 of differentiation induction (P < 0.05); however, there were no obvious differences at the other time points (P > 0.05) (Figure 6A). A western blot assay confirmed the increased expression of PPARγ on day 4 of differentiation induction (P < 0.05), although there was no obvious difference at the other time points (P > 0.05) (Figure 6B/C). These results were consistent with the results of the real-time PCR.Figure 6

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