KiSS-1, a novel human malignant melanoma metastasis-suppressor gene.

BACKGROUND Microcell-mediated transfer of chromosome 6 into human C8161 and MelJuSo melanoma cell suppresses their ability to metastasize by at least 95% without affecting their tumorigenicity. This observation demonstrates that the ability to metastasize is a phenotype distinct from tumor formation and suggests that tumorigenic cells acquire metastatic capability only after accumulating additional genetic defects. These results also imply that mutations of genes on chromosome 6 are among those late genetic changes responsible for metastatic potential. They further suggest that a melanoma metastasis-suppressor gene(s) is encoded on chromosome 6 or is regulated by genes on chromosome 6. PURPOSE Our objective was to identify the gene(s) responsible for the suppression of metastasis in chromosome 6/melanoma cell hybrids. METHODS A modified subtractive hybridization technique was used to compare the expression of messenger RNAs (mRNAs), via an analysis of complementary DNAs (cDNAs), in metastatic cells (C8161 or MelJuSo) and nonmetastatic hybrid clones (neo6/C8161 or neo6/MelJuSo). RESULTS A novel cDNA, designated KiSS-1, was isolated from malignant melanoma cells that had been suppressed for metastatic potential by the introduction of human chromosome 6. Northern blot analyses comparing mRNAs from a panel of human melanoma cells revealed that KiSS-1 mRNA expression occurred only in nonmetastatic melanoma cells. Expression of this mRNA in normal heart, brain, liver, lung, and skeletal muscle was undetectable by northern blot analysis. Weak expression was found in the kidney and pancreas, but the highest expression was observed in the placenta. The KiSS-1 cDNA encodes a predominantly hydrophilic, 164 amino acid protein with a polyproline-rich domain indicative of an SH3 ligand (binds to the homology 3 domain of the oncoprotein Src) and a putative protein kinase C-alpha phosphorylation site. Transfection of a full-length KiSS-1 cDNA into C8161 melanoma cells suppressed metastasis in an expression-dependent manner. CONCLUSIONS These data strongly suggest that KiSS-1 expression may suppress the metastatic potential of malignant melanoma cells. IMPLICATIONS KiSS-1 may be a useful marker for distinguishing metastatic melanomas from nonmetastatic melanomas.

[1]  C. Nobes,et al.  Rho, Rac, and Cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia , 1995, Cell.

[2]  A. Ridley.  Membrane ruffling and signal transduction , 1994, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  Terukatsu Sasaki,et al.  Molecular cloning of a cDNA encoding a phosphoprotein, Efs, which contains a Src homology 3 domain and associates with Fyn. , 1995, Oncogene.

[4]  S. Kottaridis,et al.  Subtraction hybridization cDNA libraries from colon carcinoma and hepatic cancer. , 1990, Genetic analysis, techniques and applications.

[5]  A. Pardee,et al.  Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. , 1993, Nucleic acids research.

[6]  G. Nicolson,et al.  Comparison of 'spontaneous' and 'experimental' metastasis using rat 13762 mammary adenocarcinoma metastatic cell clones. , 1983, Invasion & metastasis.

[7]  R. Kefford,et al.  Molecular oncogenetics of metastasis. , 1990, Molecular aspects of medicine.

[8]  R. Rickles,et al.  Phage display selection of ligand residues important for Src homology 3 domain binding specificity. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[9]  T. Pawson,et al.  SH2 and SH3 domains: From structure to function , 1992, Cell.

[10]  T Pawson,et al.  SH2 and SH3 domains , 1993, Current Biology.

[11]  I. Kjønniksen,et al.  Microenvironment revisited: Time for reappraisal of some prevailing concepts of cancer metastasis , 1994, Journal of cellular biochemistry.

[12]  Serge Vaudenay,et al.  Minding your p's and q's , 1996, ASIACRYPT.

[13]  P. Herrlich,et al.  Recent advances in the genetics of metastasis. , 1994, European journal of cancer.

[14]  E. Seftor,et al.  Characterization of a highly invasive and spontaneously metastatic human malignant melanoma cell line , 1991, International journal of cancer.

[15]  B. Mayer,et al.  SH3 Domains: Minding your p's and q's , 1995, Current Biology.

[16]  S. Schreiber,et al.  Two binding orientations for peptides to the Src SH3 domain: development of a general model for SH3-ligand interactions. , 1995, Science.

[17]  L. Liotta,et al.  Tumor invasion and metastasis: an imbalance of positive and negative regulation. , 1991, Cancer research.

[18]  E. Pick,et al.  " Peptide Walking" Is a Novel Method for Mapping Functional Domains in Proteins , 1995, The Journal of Biological Chemistry.

[19]  J. Trent,et al.  Loss of heterozygosity for loci on the long arm of chromosome 6 in human malignant melanoma. , 1991, Cancer research.

[20]  P. Meltzer,et al.  Rapid generation of region-specific genomic clones by chromosome microdissection: isolation of DNA from a region frequently deleted in malignant melanoma. , 1992, Genomics.

[21]  T Pawson,et al.  SH2 and SH3 domains: elements that control interactions of cytoplasmic signaling proteins. , 1991, Science.

[22]  B Angus,et al.  Expression of the antimetastatic gene nm23 in human breast cancer: an association with good prognosis. , 1991, Journal of the National Cancer Institute.

[23]  A. Pardee,et al.  Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. , 1992, Science.

[24]  J M Trent,et al.  Tumorigenicity in human melanoma cell lines controlled by introduction of human chromosome 6. , 1990, Science.

[25]  D. Baltimore,et al.  Proline-rich sequences that bind to Src homology 3 domains with individual specificities. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[26]  L. Liotta,et al.  Cancer metastasis and angiogenesis: An imbalance of positive and negative regulation , 1991, Cell.

[27]  D. Welch,et al.  Suppression of human melanoma metastasis by introduction of chromosome 6 may be partially due to inhibition of motility, but not to inhibition of invasion. , 1995, Biochemical and biophysical research communications.

[28]  A. V. van Kessel,et al.  Expression of nma, a novel gene, inversely correlates with the metastatic potential of human melanoma cell lines and xenografts , 1996, International journal of cancer.

[29]  L. Ellis,et al.  The implications of angiogenesis for the biology and therapy of cancer metastasis , 1994, Cell.

[30]  B. Weissman,et al.  Microcell-mediated transfer of chromosome 6 into metastatic human C8161 melanoma cells suppresses metastasis but does not inhibit tumorigenicity. , 1994, Oncogene.

[31]  G. Heijne.  A new method for predicting signal sequence cleavage sites. , 1986 .

[32]  D. Baltimore,et al.  SH2 and SH3 domains as molecular adhesives: the interactions of Crk and Abl. , 1994, Trends in biochemical sciences.

[33]  H. Bloemers,et al.  Identification of melanoma inhibitory activity and other differentially expressed messenger RNAs in human melanoma cell lines with different metastatic capacity by messenger RNA differential display. , 1995, Cancer research.

[34]  B. Seed,et al.  Isolation of cDNAs of scrapie-modulated RNAs by subtractive hybridization of a cDNA library. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[35]  C. Morelli,et al.  Induction of senescence and control of tumorigenicity in BK virus transformed mouse cells by human chromosome 6 , 1994, Genes, chromosomes & cancer.

[36]  Sam W. Lee,et al.  Positive selection of candidate tumor-suppressor genes by subtractive hybridization. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[37]  S. Barsky,et al.  The primary tumor is the primary source of metastasis in a human melanoma/SCID model. Implications for the direct autocrine and paracrine epigenetic regulation of the metastatic process , 1996, International journal of cancer.

[38]  D. Baltimore,et al.  Modular binding domains in signal transduction proteins , 1995, Cell.

[39]  Andrea Musacchio,et al.  Crystal structure of a Src-homology 3 (SH3) domain , 1992, Nature.

[40]  G. Nicolson.  Cancer progression and growth: relationship of paracrine and autocrine growth mechanisms to organ preference of metastasis. , 1993, Experimental cell research.

[41]  J. Trent,et al.  Identification of a recurring translocation site involving chromosome 6 in human malignant melanoma. , 1989, Cancer research.

[42]  L. Cantley,et al.  Oncogenes and signal transduction , 1991, Cell.

[43]  P. Fisher,et al.  Metastasis suppressed, but tumorigenicity and local invasiveness unaffected, in the human melanoma cell line MelJuSo after introduction of human chromosomes 1 or 6 , 1996, Molecular carcinogenesis.

[44]  M. Saraste,et al.  Structure and function of the SH3 domain. , 1994, Progress in biophysics and molecular biology.

[45]  J. Wright,et al.  Genetic regulation of metastatic progression. , 1990, Anticancer Research.