A role for miR-296 in the regulation of lipoapoptosis by targeting PUMA

Sophie C. Cazanave, Justin L. Mott, Nafisa A. Elmi, Steven F. Bronk, Howard Masuoka, Michael R. Charlton, Gregory J. Gores

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Saturated free fatty acids (FFA) induce hepatocyte lipoapoptosis, a key mediator of liver injury in nonalcoholic fatty liver disease (NAFLD). Lipoapoptosis involves the upregulation of the BH3-only protein PUMA, a potent pro-apoptotic protein. Given that dysregulation of hepatic microRNA expression has been observed in NAFLD, we examined the role of miRNA in regulating PUMA expression during lipotoxicity. By in silico analysis, we identified two putative binding sites for miR-296-5p within the 3′ untranslated region (UTR) of PUMA mRNA. Enforced miR-296-5p levels efficiently reduced PUMA protein expression in Huh-7 cells, while antagonism of miR-296-5p function increased PUMA cellular levels. Reporter gene assays identified PUMA 3′ UTR as a direct target of miR-296-5p. The saturated FFA, palmitate, repressed miR-296-5p expression; and Huh-7 cells were sensitized to palmitate-induced lipotoxicity by antagonism of miR-296-5p function using a targeted locked nucleic acid (LNA). Finally, miR-296-5p was reduced in liver samples from nonalcoholic steatohepatitis (NASH) patients compared with patients with simple steatosis (SS) or controls. Also miR-296-5p levels inversely varied with PUMA mRNA levels in human liver specimens. Our results implicate miR-296-5p in the regulation of PUMA expression during hepatic lipoapoptosis. We speculate that enhancement of miR-296-5p expression may represent a novel approach to minimize apoptotic damage in human fatty liver diseases.

Original languageEnglish (US)
Pages (from-to)1517-1525
Number of pages9
JournalJournal of Lipid Research
Volume52
Issue number8
DOIs
StatePublished - Aug 2011
Externally publishedYes

Fingerprint

Liver
Palmitates
3' Untranslated Regions
MicroRNAs
Nonesterified Fatty Acids
Fatty Acids
Messenger RNA
Apoptosis Regulatory Proteins
Fatty Liver
Reporter Genes
Computer Simulation
Liver Diseases
Hepatocytes
Proteins
Up-Regulation
Binding Sites
Assays
Genes
Wounds and Injuries
Non-alcoholic Fatty Liver Disease

Keywords

  • BH3-only proteins
  • Free fatty acid
  • Hepatic steatosis
  • microRNA
  • p53-upregulated mediator of apoptosis

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Cite this

Cazanave, S. C., Mott, J. L., Elmi, N. A., Bronk, S. F., Masuoka, H., Charlton, M. R., & Gores, G. J. (2011). A role for miR-296 in the regulation of lipoapoptosis by targeting PUMA. Journal of Lipid Research, 52(8), 1517-1525. https://doi.org/10.1194/jlr.M014654

A role for miR-296 in the regulation of lipoapoptosis by targeting PUMA. / Cazanave, Sophie C.; Mott, Justin L.; Elmi, Nafisa A.; Bronk, Steven F.; Masuoka, Howard; Charlton, Michael R.; Gores, Gregory J.

In: Journal of Lipid Research, Vol. 52, No. 8, 08.2011, p. 1517-1525.

Research output: Contribution to journalArticle

Cazanave, SC, Mott, JL, Elmi, NA, Bronk, SF, Masuoka, H, Charlton, MR & Gores, GJ 2011, 'A role for miR-296 in the regulation of lipoapoptosis by targeting PUMA', Journal of Lipid Research, vol. 52, no. 8, pp. 1517-1525. https://doi.org/10.1194/jlr.M014654
Cazanave, Sophie C. ; Mott, Justin L. ; Elmi, Nafisa A. ; Bronk, Steven F. ; Masuoka, Howard ; Charlton, Michael R. ; Gores, Gregory J. / A role for miR-296 in the regulation of lipoapoptosis by targeting PUMA. In: Journal of Lipid Research. 2011 ; Vol. 52, No. 8. pp. 1517-1525.
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abstract = "Saturated free fatty acids (FFA) induce hepatocyte lipoapoptosis, a key mediator of liver injury in nonalcoholic fatty liver disease (NAFLD). Lipoapoptosis involves the upregulation of the BH3-only protein PUMA, a potent pro-apoptotic protein. Given that dysregulation of hepatic microRNA expression has been observed in NAFLD, we examined the role of miRNA in regulating PUMA expression during lipotoxicity. By in silico analysis, we identified two putative binding sites for miR-296-5p within the 3′ untranslated region (UTR) of PUMA mRNA. Enforced miR-296-5p levels efficiently reduced PUMA protein expression in Huh-7 cells, while antagonism of miR-296-5p function increased PUMA cellular levels. Reporter gene assays identified PUMA 3′ UTR as a direct target of miR-296-5p. The saturated FFA, palmitate, repressed miR-296-5p expression; and Huh-7 cells were sensitized to palmitate-induced lipotoxicity by antagonism of miR-296-5p function using a targeted locked nucleic acid (LNA). Finally, miR-296-5p was reduced in liver samples from nonalcoholic steatohepatitis (NASH) patients compared with patients with simple steatosis (SS) or controls. Also miR-296-5p levels inversely varied with PUMA mRNA levels in human liver specimens. Our results implicate miR-296-5p in the regulation of PUMA expression during hepatic lipoapoptosis. We speculate that enhancement of miR-296-5p expression may represent a novel approach to minimize apoptotic damage in human fatty liver diseases.",
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AU - Gores, Gregory J.

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