Nitric oxide modulates the angiogenic phenotype of middle-T transformed endothelial cells

The role of nitric oxide (NO) in the induction of angiogenesis was evaluate d in a murine heart endothelioma cell line (H.end.FB) carrying the mT oncog ene. Two clonal derivatives of H.end.FB, H80 and H73. exhibiting different NO synthase (NOS) activities were selected and used in the study. The relat ionship among NOS activity and tumor cell behaviour (growth, and angiogenic capacity) and the molecular control of gene expression were investigated. H.end.FB and H80 on one side and H73 on the other side exhibited the highes t and lowest NOS activity, respectively. Cell growth was inversely correlat ed to the amount of NO produced by the cell lines. Conversely, in the avasc ular rabbit cornea assay. H.end.FB and H80 cells were strongly angiogenic, while H73 were poorly angiogenic, indicating that the ability of the cells to induce neovascularization was associated with the extent of NO produced. Consistently, systemic administration to rabbits of the NOS inhibitor N-w- nitro-L-arginine methyl ester (L-NAME) significantly reduced the angiogenic ity of H.end.FB cells. RT-PCR evidenced that H.end.FB expressed mRNA for TG F-beta1 and all VEGF isoforms. VEGF165 being predominantly expressed. NOS i nhibition reduced the basal expression of VEGF isoforms. while it markedly potentiated TGF-beta1 expression. These results: indicate that the endogeno us production of NO in tumor cells: can serve as an autocrine/paracrine sig nalling mechanism of progression, by controlling angiogenic factor/modulato r expression. (C) 2001 Elsevier Science Ltd. All rights reserved.