Abstract
Inflammatory and infectious conditions were simulated in cultures of ras/myc-transformed serum-free mouse embryo (ras/myc SFME) cells, using interferon-gamma (IFN-γ, 100 units/ml) and lipopolysaccharide (LPS, 0.5 μg/ml) co-treatment for 24 h, to investigate their effects on the expression of inducible nitric oxide synthase (iNOS) mRNA and the production of NO. Aminoguanidine (AG, 1 mM; an NOS inhibitor) along with IFN-γ and LPS, S-nitroso-N-acetyl-DL-penicillamine (SNAP, 100 μM; an NO donor) and/or (±)-N-[(E)-4-Ethyl-2-[(Z)-hydroxyimino]-5-nitro-3-hexene-1-yl]-3-pyridine carboxamide (NOR4, 100 μM; an NO donor), were also added to analyze the possible association of NO with matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1). Co-treatment of cells with IFN-γ and LPS increased iNOS mRNA expression, NO production, MMP-9 mRNA expression, and 105 kDa MMP-9 production. Additional treatment with the NOS inhibitor AG inhibited NO production, but did not down-regulate the expression of MMP-9 mRNA or 105 kDa MMP-9. The NO donors SNAP and NOR4 did not affect the expression of MMP-9 mRNA, 105 kDa MMP-9 or TIMP-1 mRNA. These results suggest that ras/myc SFME cells respond to infectious and inflammatory conditions and can enhance malignancy as cancer cells due to their increased levels of NO and MMP-9 production, but that NO is not directly associated with MMP-9 in these cells.
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Abbreviations
- AG:
-
Aminoguanidine
- ECM:
-
Extracellular matrix
- EGF:
-
Epidermal growth factor
- IFN-γ:
-
Interferon-gamma
- iNOS:
-
Inducible nitric oxide synthase
- IRF-1:
-
Interferon regulatory factor-1
- LPS:
-
Lipopolysaccharide
- MMP:
-
Matrix metalloproteinase
- NF-κB:
-
Nuclear factor-kappaB
- NO:
-
Nitric oxide
- NOR4:
-
(±)-N-[(E)-4-ethyl-2-[(Z)-hydroxyimino]-5-nitro-3-hexene-1-yl]-3-pyridine carboxamide
- NOS:
-
Nitric oxide synthase
- SFME:
-
Serum-free mouse embryo
- SNAP:
-
S-nitroso-N-acetyl-DL-penicillamine
- TIMP-1:
-
Tissue inhibitor of metalloproteinase-1
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Acknowledgments
We thank Dr. Shirahata of Kyushu University, Japan, for the gift of ras/myc SFME cells. This study was partly supported by a grant-in-aid from the Promotion and Mutual Aid Corporation for Private Schools of Japan.
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H. Yamaguchi and Y. Kidachi contributed equally to this work
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Yamaguchi, H., Kidachi, Y., Umetsu, H. et al. Ras/myc-transformed serum-free mouse embryo cells under simulated inflammatory and infectious conditions increase levels of nitric oxide and matrix metalloproteinase-9 without a direct association between them. Mol Cell Biochem 306, 43–51 (2007). https://doi.org/10.1007/s11010-007-9552-0
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DOI: https://doi.org/10.1007/s11010-007-9552-0