Biologic, Immunocytochemical, and Cytogenetic Characterization of Two New Human Melanoma Cell Lines: IIB‐MEL‐LES and IIB‐MEL‐IAN

Two human melanoma cell lines, derived from metastases of two patients with epithelioid malignant amelanotic melanomas, and designated IIB‐MEL‐LES and IIB‐MEL‐IAN, have been established. Both cell lines have been in continuous culture over 2 years and were propagated continuously for 85 and 75 serial passages, respectively. Morphologically, IIB‐MEL‐LES is composed predominantly of spindle shaped cells, whereas IIB‐MEL‐IAN grows as a monolayer of cuboid and stellate shaped cells with many rounded cells in suspension. Immunocytochemical studies revealed that both cell lines express S‐100 protein, vimentin, and GD₃ and GD₂ gangliosides but are negative for keratin and collagen. Both cell lines express HLA class I and HLA‐DR antigens in variable proportions. The MAGE‐1 gene is expressed only by the IIB‐MEL‐IAN cell line, as revealed by PCR analysis. Cytogenetic analysis of both cell lines revealed abnormal karyotypes; the modal chromosome numbers of IIB‐MEL‐LES and IIB‐MEL‐IAN were 48 and 81, respectively. IIB‐MEL‐LES cells presented rearrangements in chromosomes 1, 14 and X, gains in chromosomes 10,20, and 21 losses in chromosomes 15 and Y. The most frequent markers observed in IIB‐MEL‐IAN cells were 7q+, 10p+, 2p+, i(6p), 2q+, and 10q‐. Clonal gains were observed in chromosomes 12 and 21, whereas losses were seen in chromosomes 1, 2, 3, 4, 6, 7, 11, and 17. Both cell lines were capable of forming colonies in soft agar and developed tumors when transplanted into nude mice, reproducing and maintaining the characteristics of the original tumors. These cell lines and their xenografts appear to provide useful systems for studying the biology, genetics and histogenesis of human malignant melanoma and could be utilized for the development of melanoma vaccines.