Differentially expressed proteins in nitric oxide-stimulated NIH/3T3 fibroblasts: implications for inhibiting cancer development.

Abstract

PURPOSE:

Recent evidence shows that nitric oxide (NO) may exhibit both pro-cancer and anti-cancer activities. The present study aimed to determine the differentially expressed proteins in NO-treated NIH/3T3 fibroblasts in order to investigate whether NO induces proteins with pro-cancer or anti-cancer effects.

MATERIALS AND METHODS:

The cells were treated with 300 μM of an NO donor 3,3-bis-(aminoethyl)-1-hydroxy-2-oxo-1-triazene (NOC-18) for 12 h. The changed protein patterns, which were separated by two-dimensional electrophoresis using pH gradients of 4-7, were conclusively identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of the peptide digests.

RESULTS:

Seventeen differentially expressed proteins were identified in NOC-18-treated cells. Nine proteins [vinculin protein, keratin 19, ubiquitous tropomodulin, F-actin capping protein (α1 subunit), tropomyosin 3, 26S proteasome-associated pad1 homolog, T-complex protein 1 (ε subunit) N(G)-dimethylarginine dimethylaminohydrolase, and heat shock protein 90] were increased and eight proteins (heat shock protein 70, glucosidase II, lamin B1, calreticulin, nucleophosmin 1, microtubule-associated protein retinitis pigmentosa/end binding family member 1, 150 kD oxygen-regulated protein precursor, and heat shock 70-related protein albino or pale green 2) were decreased by NOC-18 in the cells. Thirteen proteins are related to the suppression of cancer cell proliferation, invasion, and metastasis while two proteins (heat shock protein 90 and N(G)-dimethylarginine dimethylaminohydrolase) are related to carcinogenesis. The functions of 150 kD oxygen-regulated protein precursor and T-complex protein 1 (ε subunit) are unknown in relation to carcinogenesis.

CONCLUSION:

Most proteins differentially expressed by NOC-18 are involved in inhibiting cancer development.

• Authors: Dong Hwi Shim, Joo Weon Lim, Hyeyoung Kim
• Journal: Yonsei medical journal (Yonsei Med J) Vol. 56 Issue 2 Pg. 563-71 (Mar 2015) ISSN: 1976-2437 [Electronic] Korea (South)
• PMID: 25684010 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• HSP70 Heat-Shock Proteins
• NOC 18
• Nitric Oxide Donors
• Nitroso Compounds
• Npm1 protein, mouse
• Proteins
• oxygen-regulated proteins
• Nucleophosmin

Topics

• Animals
• Electrophoresis, Gel, Two-Dimensional (methods)
• Fibroblasts (metabolism, pathology)
• HSP70 Heat-Shock Proteins
• Humans
• Mice
• NIH 3T3 Cells
• Neoplasms (metabolism, pathology)
• Nitric Oxide Donors
• Nitroso Compounds
• Nucleophosmin
• Proteins (analysis, metabolism)
• Proteomics (methods)
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization