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Differential activation of mitogen-activated protein kinases by γ-irradi-ation in IEC-6 cells: Role of intracellular Ca^(2+)

引用

Journal of Medical Colleges of PLA(China) 2002年第3期17卷 181-187页

作者：周舟王小华 Igisu Hideki 林远楼淑芬 Matsuoka Masato 程天民余争平 Institute of Combined Injury Third Military Medical University Chongqing 400038 China Institute of Combined Injury Third Military Medical University Chongqing 400038 China Department of Environmental Toxicology University of Occupational and Environmental Health Kitakyushu 807-85555 Japan Institute of Combined Injury Third Military Medical University Chongqing 400038 China

Objective: To explore the effects of γ-irradiation on mitogen-activated protein kinases (MAPKs) and role of intracellular calcium in this event in intestinal epithelial cell line 6 (IEC-6 cells). Methods: After cultured rat IIEC-6 cells with or without the pretreatment of intracellular Ca2+ chelator were exposed to Y-ir-radiation of 6 Gy, the total and phosphorylated MAPKs in the cells were determined with Western blotting and apoptosis was examined with flow cytometry. Activities of Extracellular signal-regulated protein kinase (ERK) and p38 MAPK were determined by using immuoprecipitation followed by Western blotting. Results: In response to γ-irradiation, phosphorylation of ERK was not significantly observed, while the levels of phosphorylated c-Jun NH2-terminal kinase (JNK) and p38 MAPK were increased in 30 min and reached the peak 2 h after exposure to 6 Gy γ-irradiation, though the cell viability was significantly lowered 12 h. On the other hand, no obvious changes were seen in the total protein levels of ERK, JNK and p38 MAPK. Chelation of intracellular Ca2+ almost completely suppressed the JNK and p38 MAPK phosphorylation induced by γ-irradia-tion, but removal of external Ca2+ had no such effect. Activation of p38 MAPK, but not of ERK, was seen to have a correlation with γ-irradiation induced apoptosis. Conclusion: The results suggest that γ-irradiation is a potent activator for JNK and p38 MAPK, and Ca2+ mobilized from intracellular stores plays an important role in the activation of MAPKs and the induction of apoptosis in IEC-6 cells.

关键词： r-irradiation extracellular signal-regulated protein kinase c-Jun NH2-terminal kinase mitogen- activated protein kinases p38 MAPK intracellular Ca2+ intestinal epithelial cell line 6

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