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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
Published on: March 16, 2017
Differentially expressed proteins in nitric oxide-stimulated NIH/3T3 fibroblasts: implications for inhibiting cancer
Dong Hwi Shim1, Joo Weon Lim2, Hyeyoung Kim3
1Department of Pharmacology, College of Medicine, Yonsei University, Seoul, Korea.
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.
Insights
Nitric oxide (NO) treatment of NIH/3T3 fibroblasts altered protein expression. Most identified proteins were associated with inhibiting cancer development, suggesting a potential anti-cancer role for NO.
Area of Science:
- Proteomics
- Cell Biology
- Cancer Research
Background:
- Nitric oxide (NO) has demonstrated dual roles in cancer, exhibiting both pro-cancer and anti-cancer activities.
- Understanding the specific protein expression changes induced by NO is crucial for elucidating its complex effects on cancer.
Purpose of the Study:
- To identify differentially expressed proteins in NIH/3T3 fibroblasts following treatment with a nitric oxide donor.
- To investigate whether NO induces proteins with pro-cancer or anti-cancer effects.
Main Methods:
- NIH/3T3 fibroblasts were treated with 300 μM of the NO donor NOC-18 for 12 hours.
- Differential protein expression was analyzed using two-dimensional electrophoresis (pH 4-7).
- Identified proteins were confirmed via MALDI-TOF MS analysis of peptide digests.
Main Results:
- Seventeen differentially expressed proteins were identified.
- Nine proteins, including vinculin and heat shock protein 90, were upregulated.
- Eight proteins, such as heat shock protein 70 and calreticulin, were downregulated.
- Thirteen proteins are linked to the suppression of cancer cell proliferation, invasion, and metastasis.
- Two proteins, heat shock protein 90 and N(G)-dimethylarginine dimethylaminohydrolase, are associated with carcinogenesis.
Conclusions:
- The majority of proteins differentially expressed by NOC-18 treatment are involved in inhibiting cancer development.
- This suggests a predominantly anti-cancer effect mediated by NO through specific protein modulation.
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