Responses of cancer cells with wild-type or tyrosine kinase domain-mutated epidermal growth factor receptor (EGFR) to EGFR-targeted therapy are linked to downregulation of hypoxia-inducible factor-1α

Background: Searching for novel molecular markers that dependably predict or indicate responses of human cancer cells to epidermal growth factor receptor (EGFR)-targeted therapy is strongly warranted. The purpose of the current study was to evaluate hypoxia-inducible factor-1 alpha (HIF-1 alpha) as a novel response marker compared with previously explored markers following treatment with an EGFR-blocking monoclonal antibody (cetuximab) and a small-molecule EGFR tyrosine kinase inhibitor (gefitinib) in a group of cancer cell lines containing wild-type or tyrosine kinase domain-mutated EGFR. Results: We found that, compared with previously studied response markers, including EGFR per se and three EGFR downstream signal molecules (ERK, Akt, and STAT3), which showed variable post- treatment changes in levels of phosphorylation and no consistent link of the changes to therapeutic responses, HIF-1 alpha showed a selective decrease in protein levels only in responsive cell lines. To demonstrate a critical role of HIF-1 alpha downregulation by EGFR-targeted treatment, we introduced a constitutively expressed HIF-1 alpha mutant (HIF-1 alpha/Delta ODD) that is resistant to cetuximab-induced downregulation in a cetuximab-responsive cell line (A431); we found that the HIF-1 alpha/Delta ODD-transfected cells remained sensitive to cetuximab-induced inhibition of Akt and ERK phosphorylation but were remarkably less responsive to cetuximab-induced growth inhibition compared with corresponding control cells. Conclusion: Our data indicates that downregulation of HIF-1 alpha is associated with positive therapeutic responses of cancer cells to EGFR-targeted therapy and suggest further investigation using HIF-1 alpha as an indicator of tumor response to EGFR-targeted therapy in preclinical studies and in the clinical setting.