Frötschl Roland, Weickardt Sandra, Staszewski Solveig, Kaufmann Gabriele, Kasper Peter
Federal Institute for Drugs and Medical Devices, Kurt-Georg-Kiesinger-Allee 3, 53175 Bonn, Germany.
Mutat Res. 2005 Aug 4;575(1-2):47-60. doi: 10.1016/j.mrfmmm.2005.03.002. Epub 2005 Apr 26.
Damage to DNA can trigger a variety of stress-related signals that alter the expression of genes associated with numerous biological pathways. In this study, we have used a cDNA microarray representing 1089 genes related to DNA damage and repair, cell cycle, transcription, metabolism and other toxicologically important cell functions to identify genes regulated in response to DNA damage in HepG2 cells induced by UV-activated chlorpromazine (CPZ). CPZ itself is not genotoxic but, upon UV irradiation with a non-genotoxic dose in the UVA range, it produces reactive free radical intermediates with DNA damaging properties. Genotoxicity in HepG2 cells was assessed concomitantly to gene expression profiling using the Comet assay. Kinetic studies were performed at a non-cytotoxic but clearly photogenotoxic concentration of CPZ (1.25 microg/ml) to characterize gene expression profiles at four different time points (3, 7, 15, 23 h) post short-term treatment. The results obtained from repeated experiments display a time-dependent expression pattern of up-regulated and repressed genes with distinct peaks in the number of differentially expressed genes at the 7 and 23 h time points. Most of the genes with altered expression belonged to the functional categories of cell cycle regulation and cell proliferation. A comparison with published expression profiles established in response to other genotoxic compounds showed low levels of concordance, which is most likely caused by the fact that extremely different testing conditions were used.
DNA损伤可触发多种与应激相关的信号,这些信号会改变与众多生物途径相关的基因表达。在本研究中,我们使用了一个cDNA微阵列,该阵列代表了1089个与DNA损伤与修复、细胞周期、转录、代谢及其他毒理学上重要的细胞功能相关的基因,以鉴定在紫外线激活的氯丙嗪(CPZ)诱导的HepG2细胞中响应DNA损伤而被调控的基因。CPZ本身无遗传毒性,但在UVA范围内用非遗传毒性剂量进行紫外线照射时,它会产生具有DNA损伤特性的活性自由基中间体。使用彗星试验在进行基因表达谱分析的同时评估HepG2细胞中的遗传毒性。在CPZ的非细胞毒性但明显具有光遗传毒性的浓度(1.25微克/毫升)下进行动力学研究,以表征短期处理后四个不同时间点(3、7、15、23小时)的基因表达谱。重复实验获得的结果显示了上调和下调基因的时间依赖性表达模式,在7小时和23小时时间点差异表达基因的数量有明显峰值。大多数表达改变的基因属于细胞周期调控和细胞增殖的功能类别。与针对其他遗传毒性化合物建立的已发表表达谱进行比较,结果显示一致性水平较低,这很可能是由于使用了极其不同的测试条件所致。