Novel MITF targets identified using a two-step DNA microarray strategy
Novel MITF targets identified using a two-step DNA microarray strategy
复制标题
DOI：
10.1111/j.1755-148x.2008.00505.x
复制
发表时间：
2008-12-01
影响因子：
4.3
通讯作者：
Steingrimsson, Eirikur
中科院分区：
文献类型：
--
作者：
Hoek, Keith S.;Schlegel, Natalie C.;Steingrimsson, Eirikur
恶性黑色素瘤是一种化疗耐药的癌症，死亡率高。最近我们在分子水平上对这种疾病的理解取得了进展，表明它与黑素细胞的发育前体有许多共同的特征，黑素细胞是皮肤和毛囊的成熟色素产生细胞。黑素细胞的发育完全依赖于小眼症相关转录因子（MITF）的作用。MITF已被证明可调节多种基因，其功能范围从色素产生到细胞周期调节、迁移和存活。然而，现有的靶点列表不足以解释MITF在黑素细胞发育和黑色素瘤进展中的作用。基因表达的DNA微阵列分析提供了一种直接的方法来识别新的靶基因，但标准的分析程序容易产生假阳性，并需要额外的实验步骤进行验证。在这里，我们介绍了一种新的策略，其中两个基于DNA微阵列的方法来识别转录因子的目标相结合的交叉验证协议，旨在帮助控制假阳性的产生。我们使用这种两步法成功地重新鉴定了13个先前记录的MITF介导的上调靶点，以及71个新靶点。这些新靶点中的许多已知与色素沉着和黑色素瘤生物学相关，并进一步强调了MITF在这些过程中的关键作用。
Malignant melanoma is a chemotherapy-resistant cancer with high mortality. Recent advances in our understanding of the disease at the molecular level have indicated that it shares many characteristics with developmental precursors to melanocytes, the mature pigment-producing cells of the skin and hair follicles. The development of melanocytes absolutely depends on the action of the microphthalmia-associated transcription factor (MITF). MITF has been shown to regulate a broad variety of genes, whose functions range from pigment production to cell-cycle regulation, migration and survival. However, the existing list of targets is not sufficient to explain the role of MITF in melanocyte development and melanoma progression. DNA microarray analysis of gene expression offers a straightforward approach to identify new target genes, but standard analytical procedures are susceptible to the generation of false positives and require additional experimental steps for validation. Here, we introduce a new strategy where two DNA microarray-based approaches for identifying transcription factor targets are combined in a cross-validation protocol designed to help control false-positive generation. We use this two-step approach to successfully re-identify thirteen previously recorded targets of MITF-mediated upregulation, as well as 71 novel targets. Many of these new targets have known relevance to pigmentation and melanoma biology, and further emphasize the critical role of MITF in these processes.