Enhancement of apoptosis in oral cancer cells by intracellular reactive oxygen amplication

利用含有约16600个基因的cDNA芯片和计算基因表达分析工具，研究了6-甲酰基蝶呤（6-FP）、α -苯基叔丁基硝基酮（PBN）和2,2-偶氮双盐酸（2-氨基丙烷）二盐酸（AAPH）自由基联合高温对人淋巴瘤U937细胞凋亡基因表达的影响。当细胞在44℃（15%凋亡水平）下处理10 min时，鉴定出39个上调基因和3个下调基因。在上调的基因中，包含凋亡和未折叠的蛋白反应相关基因。热和任何一种化学药物联合处理可提高细胞凋亡水平（约30%），并显示出化学特异性基因表达模式。此外，所选基因的表达水平通过实时定量PCR确认。本研究结果将为进一步了解不同细胞内氧化应激促进热诱导细胞凋亡的分子机制提供基础。

The effects of various free radicals derived from 6-formylpterin (6-FP), alpha-phenyl-tert-butyl nitrone (PBN) and 2,2-azobis (2-amidinopropane) dihydrochloride (AAPH) combined with hyperthermia, on gene expression in similarly enhanced apoptosis of human lymphoma U937 cells were investigated using cDNA microarrays containing approximately 16,600 genes and computational gene expression analysis tools. When the cells were treated for 10 min at 44 degrees C (15% apoptosis level), 39 up-regulated and 3 down-regulated genes were identified. In the up-regulated genes, apoptosis-and unfolded protein response-associated genes were contained. The combined treatment with heat and either chemical enhanced apoptosis level (approximately 30%) and showed a chemical-specific gene expression pattern. Furthermore, the expression levels of selected genes were confirmed by a real-time quantitative PCR. The present results will provide a basis for further understanding the molecular mechanisms in enhancement of heat-induced apoptosis by different intracellular oxidative stress.