Identification of highly expressed genes in metastasis‐suppressed chromosome 6/human malignant melanoma hybrid cells using subtractive hybridization and differential display

Microcell‐mediated transfer of chromosome 6 into human melanoma cell lines C8161 and MelJuSo suppresses metastasis by at least 95% without affecting tumorigenicity. Subtractive hybridization and differential display were used to identify the molecule(s) responsible for suppressing metastasis in neo6/melanoma (neo6/C8161 and neo6/MelJuSo) hybrids. Seven cDNA clones exhibiting quantitatively or qualitatively higher expression in neo6/melanoma hybrids were obtained. These genes fell into 2 categories: 1) transcription‐related genes (AP‐2A, HMG‐I(Y) and a novel isoform of nucleophosmin B23), which have previously been shown to regulate metastasis‐associated genes; and 2) novel genes. One of the novel genes, designated KiSS ‐1, significantly suppressed metastasis of the human malignant melanoma cell lines MelJuSo and a highly metastatic subclone of C8161, C8161cl.9, following transfection and constitutive expression. Our results illustrate the power of subtractive hybridization and differential display to identify functional metastasis‐controlling genes in human melanoma. Int. J. Cancer 71: 1035‐1044, 1997. © 1997 Wiley‐Liss Inc.