Expression analysis of Barrett's esophagus-associated high-grade dysplasia in laser capture microdissected archival tissue.
Expression analysis of Barrett's esophagus-associated high-grade dysplasia in laser capture microdissected archival tissue.
复制标题
DOI：
10.1158/1078-0432.ccr-08-0302
复制
发表时间：
2008-10-15
期刊：
影响因子：
--
通讯作者：
Resnick MB
中科院分区：
文献类型：
--
作者：
Sabo E;Meitner PA;Tavares R;Corless CL;Lauwers GY;Moss SF;Resnick MB
鉴别在非发育不良的巴雷特食管（BE）和高度发育不良（HGD）中表达的基因差异，对于提高我们对这一转变的理解有价值，并可能产生新的诊断和/或预后标志物。本研究的目的是通过对档案组织标本显微解剖的上皮细胞进行基因芯片分析，确定HGD与非发育不良BE的差异转录组。采用激光捕获显微解剖（LCM）从邻近的炎症细胞和基质细胞中分离上皮细胞。在11例患者的匹配活检中，从非发育不良BE和HGD区域提取上皮mRNA。将mRNA逆转录并应用于为福尔马林暴露组织定制的Affymetrix cDNA微阵列芯片上。对于这些基因的一个子集，通过RT-PCR和免疫组织化学证实了差异基因表达。与非发育不良BE相比，HGD中有131个基因过表达至少2.5倍，16个基因低表达至少2.5倍。在这些过表达的基因中，有几个先前被证明在be肿瘤进展中增加的基因，以及一些新的基因，如脂钙素-2、S100A9、基质金属肽酶12、丝光素1和拓扑异构酶i α。在发育不良上皮中减少的基因包括MUC5AC、三叶因子1 （TFF1）、meprin A和CD13。RT-PCR证实了28个选定基因中24个的表达变化。免疫组织化学证实，从非发育不良的BE到腺癌，整个BE相关异常增生谱中拓扑异构酶i α、S100A9和脂脂素-2的蛋白表达增加，TFF1的表达降低。这是首次在匹配患者样本中鉴定HGD与非发育不良BE中上皮基因差异表达的研究。所鉴定的基因包括几个先前与巴雷特氏相关发育不良的发病机制有关的基因，以及有待进一步研究的新候选基因。
Identifying genes differentially expressed in non-dysplastic Barrett’s esophagus (BE) from those expressed in high grade dysplasia (HGD) should be of value in improving our understanding of this transition and may yield new diagnostic and/or prognostic markers. The aim of this study was to determine the differential transcriptome of HGD compared with non-dysplastic BE through gene microarray analysis of epithelial cells microdissected from archival tissue specimens. Laser capture microdissection (LCM) was used to isolate epithelial cells from adjacent inflammatory and stromal cells. Epithelial mRNA was extracted from areas of non-dysplastic BE and HGD in matched biopsies from 11 patients. mRNA was reverse transcribed and applied on Affymetrix cDNA microarray chips customized for formalin-exposed tissue. For a subset of these genes, differential gene expression was confirmed by RT-PCR and immunohistochemistry. There were 131 genes over-expressed by at least 2.5-fold in HGD versus non-dysplastic BE and 16 genes that were under-expressed by at least 2.5-fold. Among the over-expressed genes are several previously demonstrated to be increased in the neoplastic progression of BE, as well as novel genes such as lipocalin-2, S100A9, matrix metallopeptidase 12, secernin 1 and topoisomerase IIα. Genes decreased in dysplastic epithelium include MUC5AC, trefoil factor1 (TFF1), meprin A and CD13. RT-PCR validated the changes in expression in 24 of 28 selected genes. Immunohistochemistry confirmed increased protein expression for topoisomerase IIα, S100A9 and lipocalin-2 and decreased expression of TFF1 across the spectrum of BE associated dysplasia from non-dysplastic BE through adenocarcinoma. This is the first study to identify epithelial genes differentially expressed in HGD versus non-dysplastic BE in matched patient samples. The genes identified include several previously implicated in the pathogenesis of Barrett’s-associated dysplasia and new candidates for further investigation.