Down-regulation of the Fetal Stem Cell Factor SOX17 by H33342 A MECHANISM RESPONSIBLE FOR DIFFERENTIAL GENE EXPRESSION IN BREAST CANCER SIDE POPULATION CELLS
Down-regulation of the Fetal Stem Cell Factor SOX17 by H33342 A MECHANISM RESPONSIBLE FOR DIFFERENTIAL GENE EXPRESSION IN BREAST CANCER SIDE POPULATION CELLS
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DOI：
10.1074/jbc.m109.082941
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发表时间：
2010-02-26
影响因子：
4.8
通讯作者：
Lehmann, Ulrich
中科院分区：
文献类型：
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作者：
Christgen, Matthias;Geffers, Robert;Lehmann, Ulrich
人类实体瘤含有罕见的癌旁群（SP）细胞，其排出荧光染料Hoechst 33342（H33342）并显示癌症干细胞特征。通过H33342荧光分析分离的肿瘤SP细胞的转录谱是发现肿瘤干细胞标志物和异常分化途径的新方法。利用Affyphin表达芯片和定量逆转录PCR技术，我们研究了从人乳腺癌细胞系中分离的SP和非SP（NSP）细胞之间的差异基因表达。相对于NSP细胞，乳腺癌SP中共有136个基因上调，其中之一是胎儿干细胞因子和Wnt/β-连环蛋白信号通路靶点SOX 17。引人注目的是，我们发现H33342以剂量依赖性方式下调SOX 17。在排出H33342的SP细胞中，SOX 17的下调不如保留H33342的NSP细胞中明显。因此，相对于NSP细胞，S 0X 17在癌SP中表现出10-20倍的过表达。对于其他的干相关基因，即EPC 1和SPRY 1，也获得了类似的结果。这些发现建立了一个以前未确定的基因调控的影响H33342作为一种新的机制，负责在癌症SP细胞的差异基因表达。这对未来解释癌症SP细胞具有重要意义。
Human solid tumors contain rare cancer side population (SP) cells, which expel the fluorescent dye Hoechst 33342 (H33342) and display cancer stem cell characteristics. Transcriptional profiling of cancer SP cells isolated by H33342 fluorescence analysis is a newly emerging approach to discover cancer stem cell markers and aberrant differentiation pathways. Using Affymetrix expression microarrays and quantitative reverse transcription-PCR, we investigated differential gene expression between SP and non-SP (NSP) cells isolated from human mammary carcinoma cell lines. A total of 136 genes were up-regulated in breast cancer SP relative to NSP cells, one of which was the fetal stem cell factor and Wnt/beta-catenin signaling pathway target SOX17. Strikingly, we discovered that SOX17 was down-regulated by H33342 in a dose-dependent manner. In SP cells, which expel H33342, down-regulation of SOX17 was less pronounced than in NSP cells, which retain H33342. As a result of this, SOX17 displayed a 10-20-fold overexpression in cancer SP relative to NSP cells. Similar results were obtained for further stemness-related genes, namely EPC1 and SPRY1. These findings establish a previously unidentified gene-regulatory impact of H33342 as a novel mechanism responsible for differential gene expression in cancer SP cells. This has significant implications for the future interpretation of cancer SP cells.