RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites
RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites
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DOI：
10.1073/pnas.0306988101
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发表时间：
2004-04-13
影响因子：
11.1
通讯作者：
Gordon, JI
中科院分区：
文献类型：
--
作者：
Hu, Y;Wang, T;Gordon, JI
我们以前的特点是转基因小鼠模型（CR2-TAg）的转移性前列腺癌中产生的神经内分泌（NE）细胞系。在该模型中NE分化的生物标志物在具有NE特征的常规前列腺腺癌中表达。为了进一步表征控制NE增殖、分化和存活的途径，我们建立了来自CR2-TAg前列腺肿瘤和转移的前列腺NE癌（PNEC）细胞系。在不同的通道，并作为异种移植肿瘤收获的细胞系的基因芯片分析表明，PNECs表达一致的功能，离体和体内，并与主要的CR2-TAg前列腺NE肿瘤有显着程度的相似性。PNEC表达mAsh 1，一种碱性螺旋-环-螺旋（bHLH）转录因子，对其他组织中NE细胞分化至关重要。RNA干扰敲除mAsh 1，基因芯片处理和对照细胞群的比较，以及下调基因的计算分析，确定了12个转录基序丰富的基因集。受影响的基因，包括Adcy 9，Hes 6，lapp 1，Ndrg 4，c-Myb和Mesdc 2，富含回文E-box基序，CAGCTG，表明它是一个生理相关的mAsh 1结合位点。c-Myb结合位点的富集和mAsh 1 RNA干扰下调c-Myb的发现表明mAsh 1和c-Myb处于相同的信号通路中。我们的数据表明，mAsh 1负调控细胞周期（例如，通过增强的Cdkn 2d，Bub 1表达），促进分化（例如，通过对cAMP的影响），并通过抑制细胞凋亡来增强存活。PNEC细胞系通常可用于NE细胞增殖、分化和肿瘤发生调节的遗传学和/或药理学研究。
We have previously characterized a transgenic mouse model (CR2-TAg) of metastatic prostate cancer arising in the neuroendocrine (NE) cell lineage. Biomarkers of NE differentiation in this model are expressed in conventional adenocarcinoma of the prostate with NE features. To further characterize the pathways that control NE proliferation, differentiation, and survival, we established prostate NE cancer (PNEC) cell lines from CR2-TAg prostate tumors and metastases. GeneChip analyses of cell lines harvested at different passages, and as xenografted tumors, indicated that PNECs express consistent features ex vivo and in vivo and share a remarkable degree of similarity with primary CR2-TAg prostate NE tumors. PNECs express mAsh1, a basic helix-loop- helix (bHLH) transcription factor essential for NE cell differentiation in other tissues. RNA interference knockdown of mAsh1, GeneChip comparisons of treated and control cell populations, and a computational analysis of down-regulated genes identified 12 transcriptional motifs enriched in the gene set. Affected genes, including Adcy9, Hes6, lapp1, Ndrg4, c-Myb, and Mesdc2, are enriched for a palindromic E-box motif, CAGCTG, indicating that it is a physiologically relevant mAsh1 binding site. The enrichment of a c-Myb binding site and the finding that c-Myb is down-regulated by mAsh1 RNA interference suggest that mAsh1 and c-Myb are in the same signaling pathway. Our data indicate that mAsh1 negatively regulates the cell cycle (e.g., via enhanced Cdkn2d, Bub1 expression), promotes differentiation (e.g., through effects on cAMP), and enhances survival by inhibiting apoptosis. PNEC cell lines should be generally useful for genetic and/or pharmacologic studies of the regulation of NE cell proliferation, differentiation, and tumorigenesis.