Expression profiling of microdissected pancreatic adenocarcinomas
Expression profiling of microdissected pancreatic adenocarcinomas
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DOI：
10.1038/sj.onc.1205570
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发表时间：
2002-07-04
期刊：
影响因子：
8
通讯作者：
Lemoine, NR
中科院分区：
文献类型：
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作者：
Crnogorac-Jurcevic, T;Efthimiou, E;Lemoine, NR
胰腺导管腺癌的特征是在致密促结缔组织增生性基质中嵌入少量肿瘤细胞。因此，进行激光捕获显微切割以获得正常和肿瘤性导管细胞的同质群体。利用人类 cDNA 阵列对这些样本进行基因表达的比较研究。鉴定出多种失调基因，包括细胞周期和生长调节因子、侵袭调节因子、信号传导和发育分子。除了已经发现在胰腺癌中过度表达的基因（例如 TIMP1、MMP7、CD59、rhoC 和 NDKA）之外，我们还提供证据表明以前未在该肿瘤类型中报道过的基因。其中包括过表达的基因 ABL2、Notch4 和 SOD1，以及 XRCC1（一种 DNA 修复基因，其转录本被发现下调）。采用定量实时 RT-PCR (QRT-PCR) 来确认 ABL2、Notch4 和 SOD1 的差异表达，并使用免疫组织化学分析来验证使用定制的胰腺组织阵列的 XRCC1 蛋白表达的降低。因此，将纯胰腺细胞群的微阵列衍生基因表达谱、QRT-PCR 和胰腺特异性组织阵列结合起来，在阐明这种高度恶性肿瘤类型的分子病理学方面提供了丰富的信息。
Pancreatic ductal adenocarcinoma is characterized by a paucity of neoplastic cells embedded in a densely desmoplastic stroma. Therefore, laser capture microdissection was performed to obtain homogeneous populations of normal and neoplastic ductal cells. These were subjected to a comparative study of gene expression utilizing human cDNA arrays. A variety of dysregulated genes were identified, comprising cell cycle and growth regulators, invasion regulators, signalling and developmental molecules. In addition to genes already found to be overexpressed in pancreatic cancer, such as TIMP1, MMP7, CD59, rhoC and NDKA, we present evidence to implicate genes which have not previously been reported in this tumour type. These include the overexpressed genes ABL2, Notch4 and SOD1, as well as XRCC1, a DNA repair gene whose transcript was found downregulated. Quantitative real-time RT-PCR (QRT-PCR) was employed to confirm differential expression of ABL2, Notch4 and SOD1 and immunohistochemical analysis was used to verify decreased protein expression of XRCC1 using a custom-built pancreatic tissue array. Combining microarray-derived gene expression profiles of pure pancreatic cell populations, QRT-PCR and pancreas-specific tissue arrays therefore proved to be highly informative in elucidating the molecular pathology of this highly malignant tumour type.