Expression of the Elm1 gene, a novel gene of the CCN (connective tissue growth factor, Cyr61/Cef10, and neuroblastoma overexpressed gene) family, suppresses In vivo tumor growth and metastasis of K-1735 murine melanoma cells.
We previously isolated a partial cDNA fragment of a novel gene, Elm1 (expressed in low-metastatic cells), that is expressed in low-metastatic but not in high-metastatic K-1735 mouse melanoma cells. Here we determined the full-length cDNA structure of Elm1 and investigated the effect of Elm1 expression on growth and metastatic potential of K-1735 cells. The Elm1 gene encodes a predicted protein of 367 amino acids showing approximately 40% amino acid identity with the CCN (connective tissue growth factor [CTGF], Cyr61/Cef10, neuroblastoma overexpressed gene [Nov]) family proteins, which consist of secreted cysteine-rich proteins with growth regulatory functions. Elm1 is also a cysteine-rich protein and contains a signal peptide and four domains conserved in the CCN family proteins. Elm1 was highly conserved, expressed ubiquitously in diverse organs, and mapped to mouse chromosome 15. High-metastatic K-1735 M-2 cells, which did not express Elm1, were transfected with an Elm1 expression vector, and several stable clones with Elm1 expression were established. The in vivo growth rates of cells expressing a high level of Elm1 were remarkably slower than those of cells expressing a low level of Elm1. Metastatic potential of transfectants was reduced in proportion to the level of Elm1 expression. Thus, Elm1 is a novel gene of CCN family that can suppress the in vivo growth and metastatic potential of K-1735 mouse melanoma cells.
Hashimoto
Y
Y
Biology Division, National Cancer Center Research Institute, 1-1, Tsukiji 5-chome, Chuo-ku, Tokyo 104, Japan.
Shindo-Okada
N
N
Tani
M
M
Nagamachi
Y
Y
Takeuchi
K
K
Shiroishi
T
T
Toma
H
H
Yokota
J
J
D000818:Animals; D015139:Blotting, Southern; D055511:CCN Intercellular Signaling Proteins; D002455:Cell Division / Q000235:genetics*; D003001:Cloning, Molecular; D055513:Connective Tissue Growth Factor; D055515:Cysteine-Rich Protein 61; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D016153:Genes, Suppressor / Q000235:genetics*; D008040:Genetic Linkage / Q000235:genetics; D006133:Growth Substances / Q000737:chemistry; D017874:Immediate-Early Proteins / Q000737:chemistry; D036341:Intercellular Signaling Peptides and Proteins; D008546:Melanoma, Experimental / Q000378:metabolism*; D051379:Mice; D009362:Neoplasm Metastasis / Q000235:genetics*; D055514:Nephroblastoma Overexpressed Protein; D015513:Oncogene Proteins; D009856:Oncogene Proteins, Viral / Q000737:chemistry; D011518:Proto-Oncogene Proteins / Q000737:chemistry; D012313:RNA / Q000032:analysis; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014162:Transfection / Q000235:genetics; D014407:Tumor Cells, Cultured
D055511:CCN Intercellular Signaling Proteins; C491850:CCN1 protein, mouse; C526318:CCN2 protein, mouse; C492067:CCN4 protein, mouse; C526320:Ccn3 protein, mouse; D055515:Cysteine-Rich Protein 61; D006133:Growth Substances; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D055514:Nephroblastoma Overexpressed Protein; D015513:Oncogene Proteins; D009856:Oncogene Proteins, Viral; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D055513:Connective Tissue Growth Factor; D012313:RNA