Excessive production of nitric oxide in rat solid tumor and its implication in rapid tumor growth

Koichi Doi, Takaaki Akaike, Hidechika Horie, Youichiro Noguchi, Shigemoto Fujii, Toru Beppu, Michio Ogawa, Hiroshi Maeda

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159 Citations (Scopus)


BACKGROUND. Rapid tumor growth is caused by angiogenesis factors, growth factors, etc. We previously reported a possible connection between nitric oxide (NO) and enhanced vascular permeability in solid tumor. In the present experiment, the role of NO in solid tumor pathology was further investigated in animal tumor. METHODS. To identify NO formed in solid tumor (AH136B) implanted in the feet of rats, electron paramagnetic resonance (EPR) spectroscopy was performed directly on the frozen tumor tissue at 110K by measuring endogenous nitrosyl iron-sulfur complexes, and by using exogenously added NO capturing agents, i.e., diethyldithiocarbamate (DETC)-Fe2+ and N- (dithiocarboxy)sarcosine (DTCS)-Fe2+ complexes. Induction of inducible isoform of nosymthase iNOS mRNA was examined with reverse transcriptase polymerase chain reaction (RT-PCR) combined with Southern blot analysis. In addition, vascular permeability was assessed by measuring extravasation of 51Cr labeled bovine serum albumin in solid tumor. RESULTS. Strong EPR signals from NO adducts of DETC-Fe2+ and DTCS-Fe2+ as well as strong signals from NO-hemoglobin and dinitrosyl iron sulfur complex were generated by tumor. The signal height of NO-(DTCS)2-Fe2+ observed in AH136B solid tumor was increased as the tumor gained up to 1.75 g. Induction of iNOS mRNA expression was confirmed by the above methods. Enhanced vascular permeability was suppressed by NOS inhibitors N(ω)-monomethyl-L-arginine or S- methylisothiourea sulfate and augmented with administration of L-arginine. CONCLUSIONS. Excessive NO production by iNOS in solid tumor was identified unequivocally by EPR spectroscopy. NO formed in solid tumor can be involved in enhanced vascular permeability and increased blood flow, and hence sustain tumor growth.

Original languageEnglish
Pages (from-to)1598-1604
Number of pages7
Issue number8 SUPPL.
Publication statusPublished - 1996 Apr 15


  • dithiocarbamate iron complex
  • electron paramagnetic resonance
  • enhanced vascular permeability
  • nitric oxide
  • nitric oxide synthase
  • nitrosyl iron complex
  • solid tumor pathology


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