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Effect of nitric oxide on microRNA-155 expression in human hepatic epithelial cells

  • Original Research Paper
  • Published:
Inflammation Research Aims and scope Submit manuscript

Abstract

Objective

Nitric oxide (NO) is a signaling molecule and regulator of immunity and inflammation. MicroRNAs (miRNAs) regulate gene transcription and are involved in inflammatory processes and cancer. This study sought to determine if NO activity affects miRNA expression.

Methods

Human liver epithelial (HepG2) cells were treated with the NO-releasing S-nitroso-N-acetylpenicillamine (SNAP) 100 μM for 4 h and subjected to microarray analysis. To examine the underlying mechanisms, cells were exposed to cGMP analog 8-bromo-cGMP, protein kinase inhibitor Rp-*-Br-PET-cGMPS (Rp-PET), or nitric synthase inhibitor l-NAME and evaluated with RT-PCR.

Results

MiR-155 was the only miRNA of the 887 arrayed that showed a change in expression after SNAP treatment. Incubation of the cells with 8-bromo-cGMP increased miR-155 expression 4.0 ± 0.7-fold (p < 0.05); Rp-PET before SNAP had a dual, concentration-dependent effect. SNAP treatment induced a 3.1 ± 0.7-fold change in miRNA-155 expression, Rp-PET 25 μM, a 7.3 ± 2.2-fold change, and Rp-PET 100 μM, a 0.79 ± 0.09-fold change (SNAP vs SNAP + Rp-PET, p < 0.05). In unstimulated cells, Rp-PET or l-NAME treatment increased miR-155 expression by 3.5 ± 0.7-fold and 5.6 ± 2.2-fold, respectively (p < 0.05).

Conclusion

In HepG2 cells, exogenous NO increases miR-155 expression, but endogenous basal NO inhibits it. Both effects are mediated via cGMP/PKG signaling. The upregulation of miR-155 by NO provides a new link between NO, inflammation, and cancer.

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Abbreviations

NO:

Nitric oxide

miRNA:

MicroRNA

NOS:

Nitric oxide synthase

eNOS:

Endothelial NOS

iNOS:

Inducible NOS

HepG2:

Hepatic epithelial (cells)

IL-8:

Interleukin-8

SNAP:

S-nitroso-N-acetylpenicillamine

SNP:

Sodium nitroprusside

sGC:

Soluble guanylyl cyclase

cGMP:

Cyclic guanosine monophosphate

PKG:

Protein kinase G

Rp-PET:

Rp-8-Bromo-beta-phenyl-1,N 2-ethenoguanosine 3′5′-cyclic monophosphorothioate sodium salt (Rp-*-Br-PET-cGMPS)

l-NAME:

N G-nitro-l-arginine methyl ester

Poly I:C:

Polyriboinosinic-polyribocytidylic acid

MAPK:

Mitogen-activated protein kinase

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Acknowledgments

We thank Dr. Eran Hornstein (Department of Molecular Genetics, Weizmann Institute of Science) for his help with the microarray analysis, and for critical review of the manuscript. We also thank Dr. Amiram Ravid (Felsenstein Medical Research Center) for his helpful comments.

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Authors and Affiliations

  1. Infectious Diseases Research Laboratory, Felsenstein Medical Research Center, Beilinson Campus, 49100, Petach Tikva, Israel

    Yael Yuhas, Eva Berent & Shai Ashkenazi

  2. Department of Pediatrics A, Schneider Children’s Medical Center, Petach Tikva, Israel

    Shai Ashkenazi

  3. Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

    Yael Yuhas & Shai Ashkenazi

Authors
  1. Yael Yuhas
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  2. Eva Berent
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  3. Shai Ashkenazi
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Correspondence to Yael Yuhas.

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Responsible Editor: Graham R. Wallace.

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Yuhas, Y., Berent, E. & Ashkenazi, S. Effect of nitric oxide on microRNA-155 expression in human hepatic epithelial cells. Inflamm. Res. 63, 591–596 (2014). https://doi.org/10.1007/s00011-014-0730-8

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  • DOI: https://doi.org/10.1007/s00011-014-0730-8

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