Gene expression profiling of microdissected pancreatic ductal carcinomas using high-density DNA microarrays
Gene expression profiling of microdissected pancreatic ductal carcinomas using high-density DNA microarrays
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DOI：
10.1593/neo.04295
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发表时间：
2004-09-01
期刊：
影响因子：
4.8
通讯作者：
Saeger, HD
中科院分区：
文献类型：
--
作者：
Grützmann, R;Pilarsky, C;Saeger, HD
胰腺导管腺癌（PDAC）仍然是恶性相关死亡的重要原因，也是第八大最常见癌症，总体5年相对生存率最低。为了确定新的分子标记物和新的治疗方案的候选者，我们使用Affytek U133基因芯片组研究了来自11个正常胰腺导管、14个PDAC样本和4个充分表征的胰腺癌细胞系的显微切割细胞的基因表达谱。从显微切割的样品和细胞系中提取RNA，扩增，并使用重复的体外转录方案进行标记。差异表达基因采用微阵列显著性分析程序进行鉴定。共发现616个差异表达基因。其中，140个也被其他人在PDAC中鉴定，如Galectin-1，Galectin-3和MT-SP2。我们验证了几个基因的差异表达（例如，CENPF、MCM 2、MCM 7、RAMP、IRAK 1和PTTG 1）在PDAC中的表达。我们提出了一个全基因组表达研究的显微组织从PDAC，从显微正常导管胰腺细胞和胰腺癌细胞系使用高密度微阵列。在基因组中，我们发现了新的差异表达基因，这些基因以前与PDAC的发病机制无关。
Pancreatic ductal adenocarcinoma (PDAC) remains an important cause of malignancy-related death and is the eighth most common cancer with the lowest overall 5-year relative survival rate. To identify new molecular markers and candidates for new therapeutic regimens, we investigated the gene expression profile of microdissected cells from 11 normal pancreatic ducts, 14 samples of PDAC, and 4 well-characterized pancreatic cancer cell lines using the Affymetrix U133 GeneChip set. RNA was extracted from microdissected samples and cell lines, amplified, and labeled using a repetitive in vitro transcription protocol. Differentially expressed genes were identified using the significance analysis of microarrays program. We found 616 differentially expressed genes. Within these, 140 were also identified in PDAC by others, such as Galectin-1, Galectin-3, and MT-SP2. We validated the differential expression of several genes (e.g., CENPF, MCM2, MCM7, RAMP, IRAK1, and PTTG1) in PDAC by immunohistochemistry and reverse transcription polymerase chain reaction. We present a whole genome expression study of microdissected tissues from PDAC, from microdissected normal ductal pancreatic cells and pancreatic cancer cell lines using high-density microarrays. Within the panel of genes, we identified novel differentially expressed genes, which have not been associated with the pathogenesis of PDAC before.