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Shiga toxin 1 and 2 induce apoptosis in the amniotic cell line WISH
Kazuaki Yoshimura1, Akihide Tanimoto, Tetsuya Abe
1Department of Obstetrics and Gynecology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan. yoppy@med.uoeh-u.ac.jp
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 cells 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.
Insights
Shiga toxins (Stx) 1 and 2 directly damage human amniotic cells (WISH cells) in vitro, causing cell death through apoptosis. These toxins are equally toxic to WISH and Vero cells.
Area of Science:
- Cell Biology
- Toxicology
- Molecular Biology
Background:
- Shiga toxins (Stx) are potent protein toxins produced by certain bacteria.
- Amniotic cells play a crucial role in pregnancy and fetal development.
- Understanding the effects of Stx on amniotic cells is vital for assessing potential pregnancy complications.
Purpose of the Study:
- To investigate the in vitro toxicity of Shiga toxin (Stx) 1 and 2 on human amniotic cells.
- To compare the sensitivity of amniotic cells to Stx with a known sensitive cell line (Vero cells).
Main Methods:
- Human amniotic WISH cells and Vero cells were exposed to varying concentrations of Stx 1 and 2.
- Cell viability was assessed using Cell Counting Kit-8.
- DNA damage was analyzed via agarose gel electrophoresis.
- Morphological changes and apoptosis were evaluated using Papanicolaou staining and ELISA.
Main Results:
- Stx exposure led to a dose-dependent decrease in WISH cell viability.
- DNA laddering, indicative of apoptosis, was observed in Stx-treated WISH cells.
- Morphological alterations and increased apoptotic indices confirmed Stx-induced apoptosis.
Conclusions:
- Shiga toxins directly injure human amniotic WISH cells.
- Apoptosis is the primary mechanism of cell death induced by Stx in WISH cells.
- WISH cells exhibit sensitivity to Stx comparable to Vero cells.