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[Cancer Research 56, 4224-4228, September 15, 1996]
© 1996 American Association for Cancer Research
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Role of the Human Y Box-binding Protein YB-1 in Cellular Sensitivity to the DNA-damaging Agents Cisplatin, Mitomycin C, and Ultraviolet Light1

Takefumi Ohga2, Koji Koike, Mayumi Ono, Yoshinari Makino, Yasuharu Itagaki, Morimasa Tanimoto, Michihiko Kuwano and Kimitoshi Kohno

Department of Biochemistry, Kyushu University School of Medicine, 3-1-1 Maidashi, Fukuoka 812-82 [T. O., K. Koi., M. O., Y. M., M. K.], Sapporo Research Laboratory, Snow Brand Milk Products Co., Ltd., Sapporo 065 [Y. I., M. T.], and Department of Molecular Biology, University of Occupational and Environmental Health, Kitakyushu 807 [K. Koh.], Japan

The Y box-binding protein (YB-1) binds to DNA sequences, present in the control regions of many genes, that contain an inverted CCAAT box. The binding activity of a nuclear factor, designated MDR-NF1, to an inverted CCAAT box in the promoter of the multidrug resistance 1 (MDR1) gene has previously been shown to be increased in nuclear extracts of cells exposed to UV radiation or various anticancer agents. The MDR-NF1 cDNA has now been cloned by screening a human colon library with an active fragment of the MDR1 promoter. The amino acid sequence encoded by the cloned cDNA was identical to that of YB-1. Northern blot analysis revealed that YB-1 mRNA was present in all human tissues examined. Rabbit antibodies were generated against synthetic peptides corresponding to YB-1, and indirect immunofluorescence microscopy with these antibodies showed that the concentration of YB-1 in all cisplatin-resistant cell lines examined was higher than that in the respective drug-sensitive parental cells. Transfection of human epidermoid cancer KB cells with a YB-1 antisense construct established two cell lines with reduced concentrations of YB-1. These transfectants showed increased sensitivity to cisplatin, mitomycin C, and UV radiation but not to vincristine, doxorubicin, camptothecin, or etoposide. Thus, YB-1 may protect cells from the cytotoxic effects of agents that induce cross-linking of DNA, suggesting a novel function of this ancestor DNA-binding protein.

1 This study was supported by a Grant-in-Aid for cancer research from the Ministry of Education, Science and Culture of Japan, Fukuoka Anticancer Research Fund, Yasuda Memorial Medical Fund, and the Fukuoka 21st Century Medical Fund.

2 To whom requests for reprints should be addressed. Phone: 81-92-641-1151, Ext. 3354 or 3355; Fax: 81-92-632-4198.

Received 4/24/96. Accepted 7/17/96.




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Copyright © 1996 by the American Association for Cancer Research.