1. Thai Acanthamoeba isolate (T4) induced apoptotic death in neuroblastoma cells via the Bax-mediated pathway
- Author
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Atsadang Boonmee, Araya Dharmkrong-at Chusattayanond, Siriphan Boonsilp, Saradee Warit, and Jitra Kasisit
- Subjects
Genotype ,Molecular Sequence Data ,Acanthamoeba ,Apoptosis ,DNA laddering ,Mitochondrion ,Mice ,Neuroblastoma ,Bcl-2-associated X protein ,Cell Line, Tumor ,parasitic diseases ,Animals ,Humans ,Trophozoites ,Caspase ,bcl-2-Associated X Protein ,biology ,Sequence Analysis, DNA ,Thailand ,biology.organism_classification ,Caspase Inhibitors ,Molecular biology ,Cell biology ,Infectious Diseases ,Proto-Oncogene Proteins c-bcl-2 ,Cell culture ,Caspases ,biology.protein ,DNA fragmentation ,Parasitology - Abstract
A Thai Acanthamoeba isolate named AS recovered from a corneal scraping of a keratitis patient was genotypically determined as T4. AS trophozoites were used for studying Acanthamoeba-induced apoptosis in mouse neuroblastoma NA cells during in vitro co-cultivation. The Acanthamoeba-exposed NA cells showed signs of apoptosis including cell shrinkage, nuclear condensation and DNA fragmentation. The effect was confirmed by DNA laddering electrophoresis. Involvement of caspase enzymes and mitochondrial pro- and anti-apoptotic proteins (Bax and Bcl-2) in AS-induced apoptosis was determined. The use of Z-VAD-FMK, a pan-caspase inhibitor, significantly reduced the apoptotic effect, while Bax/Bcl-2 ratio analysis showed a significant increase in the expression of apoptotic proteins in AS-exposed NA cells. These results strongly indicated that apoptosis induced by AS trophozoites is caspase-dependent and is mediated by over-expression of pro-apoptotic proteins in the mitochondrial pathway. This is the first report on the role of Bax in mediating apoptosis induced by Acanthamoeba.
- Published
- 2010
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