Targeted disruption of one allele of the Y-box binding protein-1 (YB-1) gene in mouse embryonic stem cells and increased sensitivity to cisplatin and mitomycin C

Kotaro Shibahara; Takeshi Uchiumi; Takao Fukuda; Shinobu Kura; Yohei Tominaga; Yoshihiko Maehara; Kimitoshi Kohno; Yusaku Nakabeppu; Teruhisa Tsuzuki; Michihiko Kuwano

doi:10.1111/j.1349-7006.2004.tb03214.x

Targeted disruption of one allele of the Y-box binding protein-1 (YB-1) gene in mouse embryonic stem cells and increased sensitivity to cisplatin and mitomycin C

Cancer Sci. 2004 Apr;95(4):348-53. doi: 10.1111/j.1349-7006.2004.tb03214.x.

Authors

Kotaro Shibahara¹, Takeshi Uchiumi, Takao Fukuda, Shinobu Kura, Yohei Tominaga, Yoshihiko Maehara, Kimitoshi Kohno, Yusaku Nakabeppu, Teruhisa Tsuzuki, Michihiko Kuwano

Affiliation

¹ Department of Medical Biochemistry, Kyushu University, Fukuoka 812-8582, Japan.

Abstract

The eukaryotic Y-box binding protein-1 (YB-1) functions in various biological processes, including transcriptional and translational control, DNA repair, drug resistance, and cell proliferation. To elucidate the physiological role of the YB-1 protein, we disrupted one allele of mouse YB-1 in embryonic stem (ES) cells. Northern blot analysis revealed that YB-1(+/-) ES cells with one intact allele contain approximately one-half the amount of mRNA detected in wild-type (YB-1(+/+)) cells. We further found that the protein level of YB-1(+/-) cells was reduced to approximately 50-60% compared with that of YB-1(+/+) cells. However, no apparent growth difference was found between YB-1(+/-) and YB-1(+/+) cells. YB-1(+/-) cells showed increased sensitivity to cisplatin and mitomycin C, but not to etoposide, X-ray or UV irradiation, as compared to YB-1(+/+) cells. YB-1 may have the capacity to exert a protective role against cytotoxic effects of DNA damaging agents, and may be involved in certain aspects of drug resistance.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Alleles
Animals
Antineoplastic Agents, Alkylating / pharmacology*
CCAAT-Enhancer-Binding Proteins / deficiency
CCAAT-Enhancer-Binding Proteins / genetics
CCAAT-Enhancer-Binding Proteins / physiology*
Cells, Cultured / drug effects
Cells, Cultured / metabolism
Cells, Cultured / radiation effects
Cisplatin / pharmacology*
Colony-Forming Units Assay
DNA Damage / drug effects
DNA Damage / genetics
DNA Damage / radiation effects
Drug Resistance, Neoplasm / genetics*
Embryo, Mammalian / cytology
Etoposide / pharmacology
Gene Targeting*
Heterozygote
Mice
Mitomycin / pharmacology*
NFI Transcription Factors
RNA, Messenger / genetics
Stem Cells / drug effects*
Stem Cells / metabolism
Stem Cells / radiation effects
Transcription Factors / deficiency
Transcription Factors / genetics
Transcription Factors / physiology*
Ultraviolet Rays
Y-Box-Binding Protein 1

Substances

Antineoplastic Agents, Alkylating
CCAAT-Enhancer-Binding Proteins
NFI Transcription Factors
RNA, Messenger
Transcription Factors
Y-Box-Binding Protein 1
Mitomycin
Etoposide
Cisplatin