SHORT-TERM MOLECULAR CARCINOGENICITY ASSAYS

描述：这个项目的目的是定义一组基因 表达改变，过度表达或抑制，在治疗的早期 有致癌物质的动物。作为测量变化的第一组实验 在致癌物处理期间，B6C3F1组小鼠被处理为 7，12-二甲基苯并[α]菲、呋喃和三氯乙烯。两点后 治疗数周后，获取靶组织，提取mRNAs，放射性标记 制备探针并与膜上的小鼠Atlas(TM)阵列杂交(Clontech)。 处理组织的588个序列上的表达模式为 与从未经处理的对照动物中提取的RNA进行比较。意义重大 观察到处理组和对照组之间的表达差异。在……里面 以确定这些差异是致癌物质的结果 治疗而不是简单地对所使用的化合物产生毒性，对异构体 一种是致癌的，另一种是有毒的，将用于治疗动物。这个 将测量这两种化合物的基因表达的变化，以确定 对Apri致癌物的特异性反应。这一分析将是 在至少两对化合物的治疗14天和90天时进行。 将使用小鼠Atlas(TM)阵列进行表达变化的分析 (目前1,176个序列)在玻璃幻灯片上。荧光探针将被 两种不同标记物的制备及两种探针的比较分析 是在同一张幻灯片上进行的。这些分析将扩展到诱发的肿瘤 通过使用的化合物可以获得肿瘤。基因数据库 表情变化将在第二阶段通过测试一种 大量已知致癌物改变这两个已知基因的基因表达 (例如，Atlas(TM)阵列)和随机EST。这些结果将确立 表明致癌物质的表达模式。 建议的商业应用： 这项研究的产品将是一个数据库，其中包含了 用致癌物处理过的动物的组织。这些模式的同余关系 不同类型的致癌物将建立制造 “致癌性阵列”将用于新化合物的测试。这些阵列将 首先用于为客户公司测试新化合物，作为附加数据 慢性研究，随后作为一项独立的分析。

DESCRIPTION: The purpose of this project is to define a set of genes whose expression is altered, over expression or suppression, early in treatment of an animal with a carcinogen. As a first set of experiments to measure changes in expression during carcinogen treatment, groups of B6C3F1 mice were treated with 7, 12-dimethyl benz [alpha] anthracene, furan, and trichloroethylene. After two weeks treatment, target tissues were obtained and mRNA extracted, radiolabeled probes prepared and hybridized to mouse Atlas(TM) arrays on membranes (Clontech). The expression patterns on the 588 sequences for the treated tissues were compared to those of RNA extracted from untreated control animals. Significant differences in expression between treated and control tissues were observed. In order to establish that these differences are the result of carcinogen treatment and not simply toxicity to the compounds used, pairs of isomers where one is carcinogenic and the other toxic will be used to treat animals. The changes in gene expression for the two compounds will be measured to determine the specific response to the carcinogen of the apri. This analysis will be performed at 14 and 90 days of treatment for at least two pairs of compounds. Analysis of changes in expression will be performed using mouse Atlas(TM) arrays (currently 1,176 sequences) on glass slides. Fluorescent probes will be prepared using two different labels and comparative analysis for two probes performed on the same slide. These analyses will be extended to tumors induced by the compounds used where tumors can be obtained. A database of gene expression changes will be established in phase II by testing the ability of a large number of known carcinogens to alter gene expression of both known genes (the Atlas(TM) arrays for example) and random ESTs. These results will establish expression patterns indicative of a carcinogen. PROPOSED COMMERCIAL APPLICATION: The product of this research will be a database containing the expression patterns of tissues from animals treated with carcinogens. The congruence of these patterns for different types of carcinogens will establish the genes required to manufacture a "carcinogencity array" to be used in the testing of new compounds. These arrays will be used to test new compounds for client companies first as additional data to accompany the chronic studies and subsequently as a stand alone assay.