Gene Expression Responses to Oxidative Stress

描述（由申请人提供）：许多有毒物质诱导氧化应激以及其他毒性作用，并且通常不确定这些机制如何相互作用导致病理。在本研究中，毒物多柔比星（DOX）将被用作解剖多效毒性效应的模型。DOX是一种既抑制拓扑异构酶II又引起氧化损伤的化疗药物。利用DNA微阵列技术，将首先在受到多种毒性物质（如蛋白质合成和转录抑制剂等）处理的细胞系中表征广义细胞应激的基因表达谱。然后，与topo-II抑制和氧化应激相关的谱将使用已知的毒物特异性诱导这些效应，这些谱将用于解释经DOX处理的细胞的微阵列谱。统计分析将确定DOX治疗和氧化应激的特异性变化。将研究具有不同特征（+/- p53）的乳腺基底和腔上皮细胞系，以评估细胞遗传背景对毒性的影响。DOX反应的体外数据将与现有的来自乳腺癌队列的体内基因表达文库进行比较，其中样本是在DOX治疗前后收集的。这将使研究结果转化为理解氧化应激在DOX细胞毒性中的作用的实质性贡献。

DESCRIPTION (provided by applicant): Many toxic agents induce oxidative stress along with other toxic effects, and it is often uncertain how these mechanisms interact to cause pathology. In this research, the toxicant doxorubicin (DOX) will be used as a model for dissecting pleiotropic toxic effects. DOX is a chemotherapeutic that both inhibits topoisomerase II and causes oxidative damage. Using DNA microarrays, the gene expression profile of generalized cellular stress will first be characterized in cell lines treated with diverse, well-characterized toxicants (i e protein synthesis and transcription inhibitors, etc ). Then, profiles associated with topo-II inhibition and oxidative stress will be identified using toxicants known to induce these effects specifically These profiles will be used to interpret the microarray profiles of DOX treated cells. Statistical analyses will identify changes specific to DOX treatment and oxidative stress. Basal and luminal breast epithelial cell lines with distinct characteristics (+/- p53) will be studied to assess the effect of genetic background of the cell on toxicity. The in vitro data for DOX response will be compared to an existing library of in vivo gene expression from a breast tumor cohort where samples were collected before and after DOX treatment. This will allow translation of the findings into substantial contributions to understanding of the role of oxidative stress in the cytotoxicity of DOX.