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Secreted and Cellular Polypeptide Patterns of MCF-7 Human Breast Cancer Cells following either Estrogen Stimulation or v-H-ras Transfection

Laboratory of Experimental Carcinogenesis [P. J. W., L. H., S. S. T., P. J. W.] and Medical Breast Cancer Section, Medical Branch [D. B., R. B. D., M. E. L.], National Cancer Institute, NIH, Bethesda, Maryland 20892

The polypeptide patterns of MCF-7 human breast cancer cells (MCF-7gpt) and a stably v-H-ras-transfected subclone (MCF-7ras) have been analyzed following estradiol treatment. Since both estradiol and v-H-ras transfection increase tumorigenicity of MCF-7 cells, this study was designed to ascertain if specific changes in polypeptides were common in both treatments. Separation of cellular and secreted polypeptides was accomplished by 2-dimensional polyacrylamide gel electrophoresis, and the consequent patterns were analyzed with computer assistance. Estradiol treatment of the MCF-7gpt cells reduced the number of differences found in the polypeptide patterns between MCF-7gpt and MCF-7ras. Twelve cellular polypeptides were consistently modulated by either estradiol or v-H-ras, with four polypeptides clearly affected in the same way by both treatments. Polypeptides Gchc-0845 (M_r 54,000, pI 6.9) and Gchc-0902 (M_r 52,000, pI 6.3) were suppressed by estradiol and v-H-ras, while Gchc-1240 (M_r 34,000, pI 4.4) and Gchc-1396 (M_r 23,000, pI 5.3) were induced by estradiol and v-H-ras. Sixteen secreted polypeptides were altered by at least 2-fold subsequent to estradiol treatment or v-H-ras transfection. Transfection with v-H-ras had a greater effect than estradiol, stimulating the secretion of eight polypeptides and suppressing the secretion of seven polypeptides compared to estradiol which increased secretion of five polypeptides and decreased secretion of an additional three polypeptides, respectively. Synergistic effects by estradiol and v-H-ras were noted for three polypeptides. The secretion of Gcls-175 (M_r 50,000, pI 5.7) and Gcls-320 (M_r < 14,000, pI 3.6, p-S2) was increased, while the secretion of Gcls-112 (M_r 76,000, pI 6.9) was decreased. Opposing effects of estradiol and v-H-ras were seen for seven polypeptides including the M_r 48,000 derivative of the M_r 52,000 protein (cathepsin D). These studies support the possibility that an extremely few, but specific polypeptides are regulated in association with quite diverse tumorigenic stimuli in MCF-7 human breast cancer cells.

¹ Recipient of a C. J. Martin Fellowship from the National Health and Medical Research Council of Australia. To whom requests for reprints should be addressed, at Laboratory of Experimental Carcinogenesis, National Cancer Institute, Building 37, Room 3C28, Bethesda, MD 20892.

Received 6/10/88. Revised 9/16/88. Accepted 9/27/88.

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