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Journal of the Society for Gynecologic Investigation
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Shiga Toxin 1 and 2 Induce Apoptosis in the Amniotic Cell Line WISH

Kazuaki Yoshimura, MD, PhD

Departments of Obstetrics and Gynecology, Microbiology, Pathology and Cell Biology, and Biochemistry, School of Medicine, University of Occupational and Environmental Health, Kitakyushu; Shionogi Research Laboratories, Shionogi and Co Ltd., Osaka; Department of Bacteriology, Faculty of Medicine, Kyushu University, Fukuoka, Japan yoppy{at}med.uoeh-u.ac.jp

Akihide Tanimoto, MD, PhD

Tetsuya Abe, MD, PhD

Midori Ogawa, PhD

Takashi Yutsudo, PhD

Masamichi Kashimura, MD, PhD

Shin-ichi Yoshida, MD, PhD

Departments of Obstetrics and Gynecology, Microbiology, Pathology and Cell Biology, and Biochemistry, School of Medicine, University of Occupational and Environmental Health, Kitakyushu; Shionogi Research Laboratories, Shionogi and Co Ltd., Osaka; Department of Bacteriology, Faculty of Medicine, Kyushu University, Fukuoka, Japan

Objective: The aim of this study was to evaluate the toxicity of Shiga toxin (Stx) 1 and 2 on amniotic cells in vitro.

Methods: WISH cells, which were derived from human amniotic cells, and Vero cells were cultured with or without Stxs. After 24 hours of culture, cell viability was measured by Cell Counting Kit-8, and extracted DNA was electrophoresed on a 1% agarose gel. The morphologic changes were observed by Papanicolaou staining, and the apoptotic index (percentage of apoptotic nuclei per total nuclei) was calculated. Quantification of apoptotic cells was also measured by an enzyme-linked immunosorbent assay.

Results: The viability of WISH cells decreased in proportion to the concentrations of Stxs. Cellular ladder formation was observed by DNA electrophoresis of Stx-treated WISH cells, and the typical morphologic changes were observed by Papanicolaou staining. The proportion of apoptotic cell increased in response to Stxs.

Conclusions: Stxs injured WISH cells directly and induced apoptosis in vitro. WISH cells were as sensitive as Vero cells to Stxs and cell death occurred by apoptosis.

Key Words: Shiga toxin • WISH cell • amniotic cell • apoptosis

Journal of the Society for Gynecologic Investigation, Vol. 9, No. 1, 22-26 (2002)
DOI: 10.1177/107155760200900105
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