CD4CD8 thymocytes are induced to cell death by a small dose of puromycin via ER stress |
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Authors: | Takemi Oguma Takeshi Ono Toshimitsu Kajiwara Yasushi Miyahira Yasuo Yoshihara |
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Affiliation: | a Department of Parasitology and Immunology, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan b Department of Orthopaedic Surgery, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan c Department of Global Infectious Diseases and Tropical Medicine, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan d Institute of Glycoscience, Tokai University, 1117 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan |
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Abstract: | When the CD4+CD8+ thymic lymphoma cells were treated with puromycin, we found that most of the cells died at 0.3-1 μg/ml of puromycin within 24 h. However, cell death was greatly reduced when the dose of puromycin was increased. Similar dose-pattern of cell death was observed in thymocytes and the sensitivity to puromycin was greater in CD4+CD8+ thymocytes than CD4+CD8− thymocytes. The induction of apoptosis was blocked by the protein synthesis inhibitor cycloheximide, and to some extent by transfection of Bcl-xL or Bcl-2 genes. Expression of GRP78 was up-regulated after treatment with a small dose of puromycin, and the cell death by puromycin was blocked in the presence of caspase 12 inhibitor. These results indicated that the induction of cell death by low-dose puromycin was due to endoplasmic reticulum stress. Furthermore, we found that dexamethasone, a synthetic glucocorticoid, and puromycin worked synergistically to induce cell death in thymocytes. |
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Keywords: | Puromycin CD4+CD8+ thymocytes Apoptosis ER stress |
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