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Volume 271, Number 41, Issue of October 11, 1996 pp. 25126-25130
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Identification of Histone H2A.X as a Growth Factor Secreted by an Androgen-independent Subline of Mouse Mammary Carcinoma Cells

(Received for publication, June 24, 1996)

Yoshio Watabe , Hiroaki Kuramochi , Yuzo Furuya , Nobuya Inagaki ^§ , Susumu Seino ^§ , Sadao Kimura ^¶ and Jun Shimazaki

From the  Department of Urology, the ^§ Division of Molecular Medicine, and the ^¶ Division of Cardiovascular Biology, Center for Biomedical Science, School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260, Japan

Shionogi carcinoma 115 (SC 115) cells and Chiba subline 2 (CS 2) cells are clones of an androgen-responsive mouse tumor cell line and its autonomous subline, respectively. We have shown previously that CS 2 cells produce a heparin-binding growth factor that stimulates the growth of SC 115 cells as well as the growth of themselves. In this study, a growth factor was purified from serum-free conditioned media of CS 2 cells cultured without testosterone. A heparin-binding fraction showed growth- promoting activity on SC 115 cells and BALB/3T3 cells. The amino acid sequence analysis revealed that the components were identical to histones H2A.1 and H2A.X. Since histone H2A purified from bovine thymus had almost no growth-promoting activity on SC115 cells, histone H2A.X was assumed to be a growth factor. cDNA of histone H2A.X was cloned from a library of CS 2 cells, and its sequence was confirmed. The expressed product of histone H2A.X cDNA in Escherichia coli showed remarkable stimulatory effects on growth of SC 115 cells cultured in the absence of testosterone. These results indicate that histone H2A.X is secreted from CS 2 cells cultured without testosterone and plays a role as a growth factor.

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