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Yasunobu Hashimoto^*,, Nobuko Shindo-Okada^*, Masachika Tani^*, Yasuhiro Nagamachi^*, Kaori Takeuchi^*, Toshihiko Shiroishi, Hiroshi Toma, and Jun Yokota^*

From the ^* Biology Division, National Cancer Center Research Institute, 1-1, Tsukiji 5-chome, Chuo-ku, Tokyo 104, Japan; the Department of Urology, Tokyo Women's Medical College, Kawadacho 8-chome, Shinjyuku-ku, Tokyo 162, Japan; and the Department of Mammalian Genetics Laboratory, National Institute of Genetics, 1-111, Tanida, Mishima, Shizuoka 411, Japan

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 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.

¹ Abbreviations used in this paper: bFGF, basic fibroblast growth factor; C-23, clone 23; CCN, CTGF, Cyr61/Cef10, and Nov; CT, COOH-terminal; CTGF, connective tissue growth factor; Elm, expressed in low-metastatic cells; Fisp, fibroblast-inducible secreted protein; IBP, insulin-like growth factor binding protein; mRNA, messenger RNA; MSM, Mus musculus molossinus; Nov, neuroblastoma overexpressed gene; NovM, mouse Nov; nt, nucleotide; TSP1, thrombospondin type 1 repeat; VWC, von Willebrand factor type C repeat.

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