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miR-146a targets Fos expression in human cardiac cells.

Palomer X, Capdevila-Busquets E, Botteri G, Davidson MM, Rodríguez C, Martínez-González J, Vidal F, Barroso E, Chan TO, Feldman AM, Vázquez-Carrera M - Dis Model Mech (2015)

Bottom Line: These changes correlated with a diminution in the DNA-binding activity of AP-1, the Fos-containing transcription factor complex.The specific regulation of this MMP by miR-146a was further confirmed at the secretion and enzymatic activity levels, as well as after anti-miR-mediated miR-146a inhibition.The results reported here demonstrate that Fos is a direct target of miR-146a activity and that downregulation of the Fos-AP-1 pathway by miR-146a has the capacity to inhibit MMP-9 activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Therapeutic Chemistry, IBUB (Institut de Biomedicina de la Universitat de Barcelona) and CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Faculty of Pharmacy, University of Barcelona, Diagonal 643, Barcelona E-08028, Spain.

No MeSH data available.


Related in: MedlinePlus

TNF-α upregulates miR-146a and reduces Fos expression in cardiac cells. Relative quantification of miR-146a levels in samples obtained from: (A) non-differentiated AC16 cells treated with TNF-α (100 ng/ml, 24 h); (B) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h); and (C) left ventricle tissue of transgenic TNF1.6 or control wild-type (WT) mice. Relative quantification of Fos and miR-146a expression in: (D) non-differentiated AC16 cells treated with 100 ng/ml TNF-α for 30 min to 48 h; (E) non-differentiated AC16 cells treated with 5, 10, 25, 50 and 100 ng/ml TNF-α for 24 h; and (F) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h). Graphs represent the quantification of (A-C) U6sRNA-, (D,E) 18S- or (F) APRT-normalized mRNA levels, expressed as a percentage of control (Ctrl) or wild-type (WT) samples ±s.d. *P<0.05, **P<0.01 and ***P<0.001 vs Ctrl. (G) EMSA assay showing AP-1 DNA-binding activity in non-differentiated AC16 cells treated with TNF-α. Ab, antibody; NE, nuclear extracts.
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DMM020768F1: TNF-α upregulates miR-146a and reduces Fos expression in cardiac cells. Relative quantification of miR-146a levels in samples obtained from: (A) non-differentiated AC16 cells treated with TNF-α (100 ng/ml, 24 h); (B) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h); and (C) left ventricle tissue of transgenic TNF1.6 or control wild-type (WT) mice. Relative quantification of Fos and miR-146a expression in: (D) non-differentiated AC16 cells treated with 100 ng/ml TNF-α for 30 min to 48 h; (E) non-differentiated AC16 cells treated with 5, 10, 25, 50 and 100 ng/ml TNF-α for 24 h; and (F) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h). Graphs represent the quantification of (A-C) U6sRNA-, (D,E) 18S- or (F) APRT-normalized mRNA levels, expressed as a percentage of control (Ctrl) or wild-type (WT) samples ±s.d. *P<0.05, **P<0.01 and ***P<0.001 vs Ctrl. (G) EMSA assay showing AP-1 DNA-binding activity in non-differentiated AC16 cells treated with TNF-α. Ab, antibody; NE, nuclear extracts.

Mentions: As a first approach, we assessed the effects of TNF-α on the expression of a panel of miRNAs previously related to heart disease, obesity, type 2 diabetes and inflammation. Of these, only miR-146a expression was significantly induced by TNF-α [approximately sixfold, P<0.01 vs control (Ctrl), Fig. 1A] in human cardiac AC16 cells, whereas the remaining miRNAs were not modified or simply not detected (see supplementary material Fig. S1). To further confirm these results, neonatal rat cardiomyocytes were cultured in vitro and treated with TNF-α; as shown in Fig. 1B, miR-146a was also significantly upregulated by this pro-inflammatory cytokine (1.5-fold, P<0.05 vs Ctrl). Consistent with this finding, miR-146a was also hugely stimulated in left ventricular tissue of TNF1.6 transgenic mice with cardiac-specific TNF-α overexpression [tenfold, P<0.01 vs wild type (WT), Fig. 1C].Fig. 1.


miR-146a targets Fos expression in human cardiac cells.

Palomer X, Capdevila-Busquets E, Botteri G, Davidson MM, Rodríguez C, Martínez-González J, Vidal F, Barroso E, Chan TO, Feldman AM, Vázquez-Carrera M - Dis Model Mech (2015)

TNF-α upregulates miR-146a and reduces Fos expression in cardiac cells. Relative quantification of miR-146a levels in samples obtained from: (A) non-differentiated AC16 cells treated with TNF-α (100 ng/ml, 24 h); (B) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h); and (C) left ventricle tissue of transgenic TNF1.6 or control wild-type (WT) mice. Relative quantification of Fos and miR-146a expression in: (D) non-differentiated AC16 cells treated with 100 ng/ml TNF-α for 30 min to 48 h; (E) non-differentiated AC16 cells treated with 5, 10, 25, 50 and 100 ng/ml TNF-α for 24 h; and (F) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h). Graphs represent the quantification of (A-C) U6sRNA-, (D,E) 18S- or (F) APRT-normalized mRNA levels, expressed as a percentage of control (Ctrl) or wild-type (WT) samples ±s.d. *P<0.05, **P<0.01 and ***P<0.001 vs Ctrl. (G) EMSA assay showing AP-1 DNA-binding activity in non-differentiated AC16 cells treated with TNF-α. Ab, antibody; NE, nuclear extracts.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4582106&req=5

DMM020768F1: TNF-α upregulates miR-146a and reduces Fos expression in cardiac cells. Relative quantification of miR-146a levels in samples obtained from: (A) non-differentiated AC16 cells treated with TNF-α (100 ng/ml, 24 h); (B) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h); and (C) left ventricle tissue of transgenic TNF1.6 or control wild-type (WT) mice. Relative quantification of Fos and miR-146a expression in: (D) non-differentiated AC16 cells treated with 100 ng/ml TNF-α for 30 min to 48 h; (E) non-differentiated AC16 cells treated with 5, 10, 25, 50 and 100 ng/ml TNF-α for 24 h; and (F) neonatal rat cardiomyocytes treated with TNF-α (10 ng/ml, 6 h). Graphs represent the quantification of (A-C) U6sRNA-, (D,E) 18S- or (F) APRT-normalized mRNA levels, expressed as a percentage of control (Ctrl) or wild-type (WT) samples ±s.d. *P<0.05, **P<0.01 and ***P<0.001 vs Ctrl. (G) EMSA assay showing AP-1 DNA-binding activity in non-differentiated AC16 cells treated with TNF-α. Ab, antibody; NE, nuclear extracts.
Mentions: As a first approach, we assessed the effects of TNF-α on the expression of a panel of miRNAs previously related to heart disease, obesity, type 2 diabetes and inflammation. Of these, only miR-146a expression was significantly induced by TNF-α [approximately sixfold, P<0.01 vs control (Ctrl), Fig. 1A] in human cardiac AC16 cells, whereas the remaining miRNAs were not modified or simply not detected (see supplementary material Fig. S1). To further confirm these results, neonatal rat cardiomyocytes were cultured in vitro and treated with TNF-α; as shown in Fig. 1B, miR-146a was also significantly upregulated by this pro-inflammatory cytokine (1.5-fold, P<0.05 vs Ctrl). Consistent with this finding, miR-146a was also hugely stimulated in left ventricular tissue of TNF1.6 transgenic mice with cardiac-specific TNF-α overexpression [tenfold, P<0.01 vs wild type (WT), Fig. 1C].Fig. 1.

Bottom Line: These changes correlated with a diminution in the DNA-binding activity of AP-1, the Fos-containing transcription factor complex.The specific regulation of this MMP by miR-146a was further confirmed at the secretion and enzymatic activity levels, as well as after anti-miR-mediated miR-146a inhibition.The results reported here demonstrate that Fos is a direct target of miR-146a activity and that downregulation of the Fos-AP-1 pathway by miR-146a has the capacity to inhibit MMP-9 activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Therapeutic Chemistry, IBUB (Institut de Biomedicina de la Universitat de Barcelona) and CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Faculty of Pharmacy, University of Barcelona, Diagonal 643, Barcelona E-08028, Spain.

No MeSH data available.


Related in: MedlinePlus