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[Cancer Research 61, 8887-8895, December 15, 2001]
© 2001 American Association for Cancer Research

Tumor Biology

Epidermal Growth Factor Receptor (HER1) Tyrosine Kinase Inhibitor ZD1839 (Iressa) Inhibits HER2/neu (erbB2)-overexpressing Breast Cancer Cells in Vitro and in Vivo1

Stacy L. Moulder2, F. Michael Yakes2, Senthil K. Muthuswamy3, Roberto Bianco, Jean F. Simpson and Carlos L. Arteaga4

Departments of Medicine [S. L. M., F. M. Y., R. B., C. L. A.], Cancer Biology [C. L. A.], Pathology [J. F. S.], and Vanderbilt-Ingram Cancer Center [C. L. A.], Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6307, and Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115 [S. K. M.]

Aberrrant signaling by the epidermal growth factor receptor [EGFR (HER1, erbB1)] and/or HER2/neu tyrosine kinases is present in a cohort of breast carcinomas. Because HER2 is constitutively phosphorylated in some breast tumors, we speculated that, in these cancers, transmodulation of HER2 may occur via EGFR signaling. To test this possibility, we examined the effect of EGFR-specific kinase inhibitors against the HER2-overexpressing human breast tumor lines BT-474, SKBR-3, MDA-361, and MDA-453. ZD1839 (Iressa) is an ATP-mimetic that inhibits the purified EGFR and HER2 kinases in vitro with an IC50 of 0.033 and >3.7 µM, respectively. The specificity of ZD1839 against EGFR was confirmed in Rat1 fibroblasts transfected with EGFR or HER2 chimeric receptors activated by synthetic ligands without the interference of endogenous receptors. Treatment of all breast cancer cell lines (except MDA-453) with 1 µM ZD1839 almost completely eliminated HER2 phosphorylation. In contrast, the incorporation of [{gamma}-32P]ATP in vitro onto HER2 receptors isolated from BT-474 cells was unaffected by 1 µM ZD1839. EGFR is expressed by BT-474, SKBR-3, and MDA-361 but not by MDA-453 cells, suggesting that ZD1839-mediated inhibition of the EGFR kinase explained the inhibition of HER2 phosphorylation in vivo. In SKBR-3 cells, ZD1839 exhibited a greater growth-inhibitory effect than Herceptin, a monoclonal antibody against the HER2 ectodomain. In both SKBR-3 and BT-474 cells, treatment with ZD1839 plus Herceptin induced a greater apoptotic effect than either inhibitor alone. Finally, ZD1839 completely prevented growth of BT-474 xenografts established in nude mice and enhanced the antitumor effect of Herceptin. These data imply that EGFR tyrosine kinase inhibitors will be effective against HER2-overexpressing breast tumor cells that also express EGFR and support their use in combination with HER2 antibodies, such as Herceptin, against mammary carcinomas with high levels of the HER2 proto-oncogene.




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