Modulation of heat-induced cell death in PC-3 prostate cancer cells by the antioxidant inhibitor diethyldithiocarbamate
Modulation of heat-induced cell death in PC-3 prostate cancer cells by the antioxidant inhibitor diethyldithiocarbamate
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DOI：
10.1046/j.1464-410x.2002.02810.x
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发表时间：
2002-08-01
影响因子：
4.5
通讯作者：
Terashima, M
中科院分区：
文献类型：
--
作者：
Moriyama-Gonda, N;Igawa, M;Terashima, M
目的探讨PC-3前列腺癌细胞经热疗后细胞死亡形式(细胞凋亡或坏死)、活性氧(ROS)生成、超氧化物歧化酶(SOD)活性与热休克蛋白70(HSP 70)表达水平的关系，以及热疗与抗氧化剂二乙基二硫代氨基甲酸酯(DDC)联合作用对PC-3细胞的杀伤作用。细胞增殖、ROS生成、超氧化物歧化酶(SOD)活性和细胞热休克蛋白70(HSP70)水平分别采用四唑盐细胞毒试验、荧光二氯荧光素(DCF)和硝基蓝四氮唑蓝比色法、Western印迹分析和流式细胞术检测。用碘化丙啶和异硫氰酸荧光素标记的膜联蛋白V染色检测PC-3细胞的凋亡数和坏死率，并在43℃和44℃温热处理PC-3细胞60min后，检测细胞凋亡率和坏死率。结果43℃加热60、90、120min和44℃时，细胞存活率均显著降低(P
Objective To examine the relationships between the form of cell death (apoptosis or necrosis), reactive oxygen species (ROS) generation, superoxide dismutase (SOD) activity and the level of heat-shock protein 70 (hsp 70) expression after thermotherapy of PC-3 prostate cancer cells; also assessed were the tumoricidal effects of combined treatment with both heat and the antioxidant inhibitor diethyldithiocarbamate (DDC).Materials and methods PC-3 cells were treated with thermotherapy at 42, 43 or 44degreesC for 30, 60, 90 or 120 min. Cell proliferation, ROS generation, SOD activity and cellular hsp 70 level were determined using tetrazolium-based cytotoxicity, fluorescent dichlorofluorescein (DCF) and nitroblue tetrazolium assays, Western blot analysis and flow cytometry, respectively. The apoptotic and necrotic cells were determined by staining with propidium iodide and fluorescein isothiocyanate-labelled annexin V. These variable were also measured after combined treatment of PC-3 cells with 1 mmol/L DDC and thermotherapy at 43 or 44degreesC for 60 min.Results Cell survival was significantly lower after heating cells at 43degreesC for 60, 90 and 120 min and at 44degreesC for all periods tested (P