Differentially Expressed Proteins in Nitric Oxide-Stimulated NIH/3T3 Fibroblasts: Implications for Inhibiting Cancer Development.
10.3349/ymj.2015.56.2.563
- Author:
Dong Hwi SHIM
1
;
Joo Weon LIM
;
Hyeyoung KIM
Author Information
1. Department of Pharmacology, College of Medicine, Yonsei University, Seoul, Korea. kim626@yonsei.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Nitric oxide;
proteomic analysis;
cancer;
NIH/3T3 cells
- MeSH:
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;
Proteins/analysis/*metabolism;
Proteomics/*methods;
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- From:Yonsei Medical Journal
2015;56(2):563-571
- CountryRepublic of Korea
- Language:English
-
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 microM 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 (alpha1 subunit), tropomyosin 3, 26S proteasome-associated pad1 homolog, T-complex protein 1 (epsilon 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 (epsilon subunit) are unknown in relation to carcinogenesis. CONCLUSION: Most proteins differentially expressed by NOC-18 are involved in inhibiting cancer development.
