Molecular analysis of intestinal stem cell associated genes in colorectal cancer (CRC)

一项综合的分子图谱研究将原发结直肠肿瘤与同一患者的肝转移瘤进行了比较，发现了在转移过程中差异调控的基因和途径。另一组重要的候选基因来自于对CD133阳性结直肠癌细胞中激活基因的鉴定，CD133阳性结直肠癌细胞被认为代表了富含癌症干细胞的细胞部分。本项目的目的是对这两组候选基因进行功能鉴定，以阐明它们在结直肠癌发生和转移过程中的作用。候选基因的表达将在正常和结直肠癌细胞系中进行调节，表型效应将通过应用测试细胞黏附、细胞迁移和侵袭的分析来研究。基因调控的表达和DNA甲基化分析将允许对候选基因启动的与转移相关的过程中涉及的分子路径和网络产生系统的看法。遵循癌症干细胞äS作为转移的基础细胞的概念，将使用椭球培养深入分析来自两个顶级途径的顶级基因和成员的作用。使用这个主要的体外系统和异种移植模型的工作将与子项目3和其他子项目密切合作进行。此外，我们将使用具有生物活性的真菌提取物和/或小分子筛选设施(在EMBL和DKFZ之间建立)来筛选新的抑制剂。这些抑制物在具有转移相关表型的细胞系统中的测试，将允许验证新出现的分子干扰策略，这是在这个子项目过程中开发的。

An integrated molecular profiling effort compared primary colorectal tumors with liver metastases from the same patient and revealed genes and pathways that are differentially regulated in the process of metastasis. Another set of important candidate genes was derived from the identification of genes activated in CD133-positive colorectal tumor cells, which are considered to represent a cellular fraction enriched in cancer stem cells. The aim of this project is to functionally characterize these two sets of candidate genes in order to elucidate their role in colorectal carcinogenesis and the process of metastasis. Candidate gene expression will be modulated in normal and colorectal cell lines and phenotypic effects will be investigated by applying assays which test cell adhesion, cell migration and Invasion. Expression and DNA methylation profiling upon gene modulation will allow the generation of a systematic view of the molecular pathways and networks involved in metastasis-related processes initiated by our candidate genes. Following the concept of cancer stem cells äs the founding cells of metastases, the effect of top-ranked genes and members from the two top-ranked pathways will be analyzed in depth using spheroid cultures. Work using this prime in vitro System and xenotransplant models will be performed in close collaboration with subproject 3 and other subprojects. Furthermore, we will screen for novel inhibitors using biologically active fungi extracts and/or the small molecular screening facility (established between the EMBL and the DKFZ). The testing of these inhibitors in the cellular Systems with metastasis-related phenotype, which are developed in the course of this subproject, will allow validation of newly emerging molecular interference strategies.