Identification of SMURF1 as a possible target for 7q21.3-22.1 amplification detected in a pancreatic cancer cell line by in-house array-based comparative genomic hybridization
Identification of SMURF1 as a possible target for 7q21.3-22.1 amplification detected in a pancreatic cancer cell line by in-house array-based comparative genomic hybridization
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DOI：
10.1111/j.1349-7006.2008.00779.x
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发表时间：
2008-05-01
期刊：
影响因子：
5.7
通讯作者：
Imoto, Issei
中科院分区：
文献类型：
--
作者：
Suzuki, Ayako;Shibata, Tatsuhiro;Imoto, Issei
胰腺癌（PC）细胞系为发现和功能分析驱动这种致命癌症发生和发展的基因提供了一个有用的起点。为了增加我们对胰腺癌中基因拷贝数变化的理解，并确定关键的扩增和缺失靶点，我们使用内部阵列（MCG Cancer Array-800）对24个PC细胞系进行了基于全基因组阵列的比较基因组杂交。总的来说，分析揭示了高基因组复杂性，在每个系中检测到几个拷贝数变化。在24个细胞系中的14个中观察到8个基因（克隆）的纯合缺失（log（2）比率<-2），而在7个细胞系中检测到10个基因（克隆）的高水平扩增（log（2）比率> 2）。其中，我们专注于7q22.1的高水平扩增，因为这种改变的靶基因仍然未知。通过荧光原位杂交对改变区域进行精确定位，确定位于这些区域内的基因的表达状态，并在PC细胞系中使用基因表达敲低或异位过表达方法进行功能分析，以及对PC原发性肿瘤中的候选者进行免疫组织化学分析，我们成功地鉴定了SMURF1作为7q21.3 - 22.1扩增靶具有最大的潜力。SMURF1可能通过过表达在PC中发挥促生长基因的作用，并可能成为一个很好的治疗靶点。我们的研究结果表明，基于阵列的比较基因组杂交分析结合进一步的遗传和功能检查是一种有用的方法，用于确定新的肿瘤相关基因参与这种致命疾病的发病机制。
Pancreatic cancer (PC) cell lines provide a useful starting point for the discovery and functional analysis of genes driving the genesis and progression of this lethal cancer. To increase our understanding of the gene copy number changes in pancreatic carcinomas and to identify key amplification and deletion targets, we applied genome-wide array-based comparative genomic hybridization using in-house array (MCG Cancer Array-800) to 24 PC cell lines. Overall, the analyses revealed high genomic complexity, with several copy number changes detected in each line. Homozygous deletions (log(2)ratio < -2) of eight genes (clones) were seen in 14 of the 24 cell lines, whereas high-level amplifications (log(2)ratio > 2) of 10 genes (clones) were detected in seven lines. Among them, we focused on high-level amplification at 7q22.1, because target genes for this alteration remain unknown. Through precise mapping of the altered region by fluorescence in situ hybridization, determination of the expression status of genes located within those regions, and functional analysis using knockdown of the gene expression or the ectopic overexpression approach in PC cell lines, as well as immunohistochemical analyses of candidates in primary tumors of PC, we successfully identified SMURF1 as having the greatest potential as a 7q21.3-22.1 amplification target. SMURF1 may work as a growth-promoting gene in PC through overexpression and might be a good candidate as a therapeutic target. Our results suggest that array-based comparative genomic hybridization analysis combined with further genetic and functional examinations is a useful approach for identifying novel tumor-associated genes involved in the pathogenesis of this lethal disease.