INSULIN-LIKE GROWTH-FACTOR (IGF) BINDING TO A MURINE MAMMARY EPITHELIAL-CELL LINE

The goal of this study was to relate insulin-like growth factor (IGF) binding with IGF-stimulated growth in a murine mammary epithelial (COM MA-DIMME) cell line. Affinity crosslinking with [I-125]IGF-I showed ma jor bands at 224,000 and 148,000 Mr that were ablated by the inclusion of IGF-I or -II at 130 nM. Scatchard-transformed [I-125] IGF-I bindin g data was best fit by a two-site model, with 24,000 sites possessing a Kd of 0.33 nM and 1,000 sites possessing a Kd of 8.09 nM per cell. C ompetition analysis showed ED,, values for ICE-I, -II, and insulin to be 0.90 +/- 0.03, 7.15 +/- 4.27, and 335 +/- 104 nM, respectively. Aff inity crosslinking of [I-125]ICF-II showed three major bands of 230,00 0, 148,000, and 61,000 to 58,000 Mr. Unlabeled IGF-II ablated all band s, while IGF-I and insulin ablated only the 148,000 Mr band. Competiti on analysis showed ED,, values for unlabeled IGF-I and -II to be 0.10 +/- 0.01 and 5.31 +/- 2.04 nM, respectively. In spite of the receptor affinity differences, no significant difference was noted in IGF-I and -II in capacity to stimulate cell growth. These data indicate that CO MMA-DIMME cells express IGF receptors and that both IGFs are mitogenic . (C) 1994 Wiley-Liss, Inc.