Molecular and Cellular Biochemistry

, Volume 306, Issue 1–2, pp 43–51 | Cite as

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

  • Hideaki Yamaguchi
  • Yumi Kidachi
  • Hironori Umetsu
  • Kazuo Ryoyama


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.


Interferon-gamma (IFN-γ) Lipopolysaccharide (LPS) Matrix metalloproteinase (MMP) Nitric oxide (NO) ras/myc-transformed serum-free mouse embryo (ras/myc SFME) Serum-free mouse embryo (SFME) 





Extracellular matrix


Epidermal growth factor




Inducible nitric oxide synthase


Interferon regulatory factor-1




Matrix metalloproteinase


Nuclear factor-kappaB


Nitric oxide


(±)-N-[(E)-4-ethyl-2-[(Z)-hydroxyimino]-5-nitro-3-hexene-1-yl]-3-pyridine carboxamide


Nitric oxide synthase


Serum-free mouse embryo




Tissue inhibitor of metalloproteinase-1



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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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Hideaki Yamaguchi
    • 1
    • 2
  • Yumi Kidachi
    • 1
  • Hironori Umetsu
    • 3
  • Kazuo Ryoyama
    • 1
    • 2
  1. 1.Department of Clinical Pharmacy, Faculty of Pharmaceutical SciencesAomori UniversityAomoriJapan
  2. 2.Graduate School of Environmental SciencesAomori UniversityAomoriJapan
  3. 3.Laboratory of Food Chemistry, Department of Life Sciences, Junior CollegeGifu Shotoku Gakuen UniversityGifuJapan

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