Expression of angiogenic factors vascular endothelial growth factor and interleukin-8/CXCL8 is highly responsive to ambient glutamine availability: Role of nuclear factor-κB and activating protein-1

Vascular endothelial growth factor (VEGF) and interleukin-8/CXCL8 (IL-8) are prominent pro-angiogenic and pro-metastatic proteins that represent negative prognostic factors in many types of cancer. Hypoxia is thought to be the primary environmental cause of VEGF and IL-8 expression in solid tumors. We hypothesized that a lack of nutrients other than oxygen could stimulate the expression of these factors and previously demonstrated that expression of VEGF and IL-8 is responsive to amino acid deprivation. In the present study, we examined the effect of glutamine availability on the expression of these factors as well as the role of transcription factors NF Kappa B and activating protein-1 (AP-1) in the response of TSE human breast carcinoma cells to glutamine deprivation. VEGF and IL-8 secretion and mRNA levels were dramatically induced by glutamine deprivation. mRNA stabilization contributed to this response. Glutamine deprivation increased NF Kappa B (p65/p50) and AP-1 (Fra-1/c-Jun+JunD) DNA- binding activities. Blocking NF Kappa B and AP-1 activation with curcumin as well as expression of dominant inhibitors, inhibitor of nuclear factor- Kappa B (I Kappa B) super repressor (I Kappa BM), and a mutant form of c-Fos (A-Fos) demonstrated that the activation of NF Kappa B and AP-1 transcription factors was necessary for the induction of IL-8 expression but dispensable for the induction of VEGF expression. A macro-array containing 111 NF Kappa B target genes identified a total of 17 that were up-regulated 2-fold or more in response to glutamine deprivation. These included growth regulated oncogene alpha (GRO alpha /GRO1/CXCL1), another neutrophil chemoattractant implicated in tumor angiogenesis and metastasis.