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Extraction of a Melanoma Growth-stimulatory Activity from Culture Medium Conditioned by the Hs0294 Human Melanoma Cell Line¹

Clinical Research Facility [A. R., J. S. S., R. K. C.], Division of Hematology-Oncology [D. H. L., D. W. N.], and Winship Clinic for Neoplastic Diseases [D. W. N.], Department of Medicine, Emory University School of Medicine, Atlanta, Georgia 30322

Extracts of conditioned medium (CM) from Hs0294 human malignant melanoma cells stimulate [³H]thymidine incorporation and an increase in cell number in serum-depleted Hs0294 cells. This activity is acid and heat stable, nonproteolytic, protease sensitive, contains disulfide bonds and elutes broadly from a Bio-Gel P-30 column with an approximate molecular weight range of 6,000 to 25,000. Hs0294 CM also stimulates [³H]thymidine incorporation in nonmalignant human nevus cells and normal rat kidney fibroblast cells but not in human fibroblasts. There was only limited competition with ¹²⁵I-epidermal growth factor in binding assays. Hs0294 CM extracts stimulate anchorage-independent growth in normal rat kidney fibroblast cells in soft agar but not in Hs0294 cells, nevus cells, or human fibroblasts. This second activity elutes from the Bio-Gel P-30 column in two positions with apparent molecular weights of 27,000 and 11,000.

¹ Supported by The McCandless Fund and The Woodruff Ad Hoc Fund of Emory University, Biomedical Research Fund NIH RR-05364, State of Georgia Nutrition in Cancer Contract, Austin White Charitable Trust Fund, NIH Grant RR00039, NIH Fellowship 1 F32 AM06426-01 and the Winship Clinic Research Fund. Presented in part to the American Association for Cancer Research, Washington, D. C., April 26 to 30, 1981 (13); The Ninth Cold Spring Harbor Conference on Cell Proliferation, Cold Spring Harbor, N. Y., September 1 to 6, 1981 (12); and the Federation of American Societies for Experimental Biology, New Orleans, La., April 16 to 23, 1982 (11).

² To whom requests for reprints should be addressed.

Received 8/25/81. Accepted 1/19/83.

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