Vascular endothelial growth factor receptor-1 contributes to resistance to anti - epidermal growth factor receptor drugs in human cancer cells

Abstract

Purpose:The resistance to selective EGFR inhibitors involves the activation of alternative signaling pathways, and Akt activation and VEGF induction have been described in EGFR inhibitor - resistant tumors. Combined inhibition of EGFR and other signaling proteins has become a successful therapeutic approach, stimulating the search for further determinants of resistance as basis for novel therapeutic strategies. Experimental Design: We established human cancer cell lines with various degrees of EGFR expression and sensitivity to EGFR inhibitors and analyzed signal transducers under the control of EGFR-dependent and EGFR-independent pathways. Results: Multitargeted inhibitor vandetanib (ZD6474) inhibited the growth and the phosphorylation of Akt and its effector p70S6 kinase in both wild-type and EGFR inhibitor-resistant human colon, prostate, and breast cancer cells. We found that the resistant cell lines exhibit, as common feature, VEGFR-1/Flt-1 overexpression, increased secretion of VEGF and placental growth factor, and augmented migration capabilities and that vandetanib is able to antagonize them. Accordingly, a new kinase assay revealed that in addition to VEGF receptor (VEGFR) -2, RET, and EGFR, vandetanib efficiently inhibits also VEGFR-1. The contribution of VEGFR-1 to the resistant phenotype was further supported by the demonstration that VEGFR-1 silencing in resistant cells restored sensitivity to anti-EGFR drugs and impaired migration capabilities,whereas exogenous VEGFR-1 overexpression in wild-type cells conferred resistance to these agents. Conclusions: This study shows that VEGFR-1 contributes to anti-EGFR drug resistance in different human cancer cells. Moreover, vandetanib inhibits VEGFR-1 activation, cell proliferation, and migration, suggesting its potential utility in patients resistant to EGFR inhibitors. © 2008 American Association for Cancer Research. We report several findings that may have relevant clinical and therapeutic implications. First, using a panel of tumor cells of different types and with different degrees of sensitivity or resistance to epidermal growth factor receptor (EGFR) inhibitors, we show that resistant tumors share the following common features: vascular endothelial growth factor receptor (VEGFR) -1/Flt-1 overexpression and Akt activation, increased secretion of VEGF and placental growth factor, and augmented migration capabilities. We also provide mechanistic evidence of the correlation between VEGFR-1 activity and EGFR drug resistance. These data imply that detection of VEGFR-1 on tumor cells may indicate their increased ability to survive and invade and to escape the inhibition by EGFR inhibitors used in clinical practice, such as cetuximab and gefitinib. Another finding concerns the small-molecule vandetanib/ZD6474, whose mechanisms of action have been documented by us in several previous studies. We here show that vandetanib is able to inhibit VEGFR-1 kinase and EGFR drug-resistant tumors. On these bases, we have remeasured and reported a new kinase inhibition profile for this drug. Because vandetanib is under investigation in several clinical studies, these data may be important for its clinical development. © 2008 American Association for Cancer Research.