Prediction of nitric oxide concentrations in melanomas

Abstract

The presence of iNOS and nitrotyrosine in cutaneous melanomas has been correlated with poor survival rates of patients, suggesting that NO plays a role in the tumor pathophysiology. However, the concentrations of NO that melanoma cells are exposed to in vivo have been unknown. To provide cell kinetic data for use in predicting those concentrations, synthesis and consumption of NO was examined in A375 melanoma cells. Nitric oxide synthesis was undetectable. The rate of intracellular NO consumption was determined by continuous monitoring of NO concentrations following injection of NO solutions in a closed chamber. After correcting for autoxidation and consumption from media-generated O 2 - , the rate constant obtained for cellular consumption was 7.1 ± 1.1 s −1. This information was combined with previous data on macrophage NO kinetics to develop a mathematical model to predict NO levels in cutaneous melanomas. Synthesis of NO by macrophages in the stroma was found to give a maximum concentration at the tumor periphery of 0.2 μM. Because of the high rates of cellular consumption, the elevation in NO concentration is predicted to be very localized, approximately 90% of the concentration decay occurring within 30 μm of the tumor edge. High NO concentrations at the periphery of a melanoma may contribute to metastasis by stimulating cell proliferation, inhibiting apoptosis, or acting as a lymphangiogenic factor.