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Epidermal Growth Factor Receptors and Effect of Epidermal Growth Factor on Growth of Human Breast Cancer Cells in Long-Term Tissue Culture¹

Department of Physiology, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada, R3E 0W3

Epidermal growth factor (EGF) may be important in regulating the proliferation of mammary epithelial cells. In the present study, we examined EGF binding and effect on growth in nine human mammary cell lines. The T-47D, MCF-7, SK-Br-3, AlAb 496, BT-20, and BT-474 tumor cell lines and a cell line (HBL-100) derived from milk exhibited EGF binding; both high (K_a 10¹⁰ M^-1)- and low (K_a 10⁹ M^-1)-affinity sites were detected. The total number of EGF receptors per cell of different cell lines varied from 1.6 x 10³ sites/cell (for AlAb 496) to 1.5 x 10⁶ sites/cell (for BT-20). The two floating cell lines, DU4475 and Lev III, had no detectable EGF binding. Effect of EGF on growth was studied by monitoring cell number and the incorporation of [³H]thymidine into DNA of cells maintained in Dulbecco's modified Eagle's medium supplemented with 0.1% fetal bovine serum. Using these procedures, only T-47D cells were stimulated by EGF at low concentrations (0.1 to 1 ng/ml). At concentrations higher than 10 ng/ml, EGF was inhibitory to varying degrees in most cell lines that contained EGF receptors. The growth of the two floating cell lines that had no detectable EGF binding was unaffected by EGF. Our results show that EGF receptors are not present in all human breast cancer cell lines. There is no apparent correlation between EGF binding and its mitogenic activity in cell lines with EGF receptors. EGF may have biological roles in human breast cancer other than growth regulation.

¹ Supported by the Medical Research Council of Canada and the National Cancer Institute of Canada. Portions of this work were reported at the Twenty-First Annual Meeting of the American Society for Cell Biology held in Anaheim, Calif., November 9 to 13, 1981 (14).

² To whom requests for reprints should be addressed.

Received 2/ 4/82. Accepted 7/26/82.

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