Establishment of the epithelial-specific transcriptome of normal and malignant human breast cells based on MPSS and array expression data.
Establishment of the epithelial-specific transcriptome of normal and malignant human breast cells based on MPSS and array expression data.
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DOI：
10.1186/bcr1604
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
O'Hare MJ
中科院分区：
文献类型：
--
作者：
Grigoriadis A;Mackay A;Reis-Filho JS;Steele D;Iseli C;Stevenson BJ;Jongeneel CV;Valgeirsson H;Fenwick K;Iravani M;Leao M;Simpson AJ;Strausberg RL;Jat PS;Ashworth A;Neville AM;O'Hare MJ
各种基因芯片和测序技术已被广泛用于检测乳腺癌恶性上皮细胞的分子变化。然而，这种基因表达研究，以确定肿瘤细胞中的标志物和目标，受到损害的实体乳腺肿瘤的细胞异质性和缺乏适当的对应物代表正常的乳腺上皮细胞。采用免疫磁性分离方法分离了原发性乳腺癌的恶性肿瘤上皮细胞和正常人乳腺组织的管腔上皮细胞和肌上皮细胞。使用大规模平行签名测序（MPSS）和四种不同的全基因组微阵列平台，对来自这些细胞类型的高度富集制剂的RNA进行表达谱分析。差异肿瘤上皮转录组的功能相关转录物用于基因集富集分析，以鉴定管腔型和肌上皮型基因的富集。在组织微阵列上进行了少量基因的临床病理学验证。MPSS在正常管腔细胞池和基本上富含上皮细胞的原发性肿瘤之间鉴定了6，553个差异表达基因，其中98%被代表，60%被微阵列分析证实。仅通过微阵列技术检测到的这两个样品之间的显著表达水平变化由4，149个转录物显示，导致8，051个基因的组合差异肿瘤上皮转录组。微阵列基因签名确定了907和955个转录本的综合列表，其表达分别在腔上皮细胞和肌上皮细胞之间不同。功能注释和基因集富集分析突出了一组与骨骼发育相关的基因，这些基因与肌上皮/基底细胞相关，并在肿瘤样品中上调。在乳腺癌组织微阵列上，对这一类别中最高度过表达的基因之一编码骨膜蛋白进行了化学分析，其在肿瘤细胞中的表达与一组预后不良的雌激素受体阳性肿瘤的预后不良相关。使用高度富集的细胞群结合多平台基因表达谱研究，建立了正常和恶性乳腺组织之间分子变化的综合分析。本研究为乳腺癌的诊断、预后和治疗提供了新的和潜在的重要靶点。
Diverse microarray and sequencing technologies have been widely used to characterise the molecular changes in malignant epithelial cells in breast cancers. Such gene expression studies to identify markers and targets in tumour cells are, however, compromised by the cellular heterogeneity of solid breast tumours and by the lack of appropriate counterparts representing normal breast epithelial cells. Malignant neoplastic epithelial cells from primary breast cancers and luminal and myoepithelial cells isolated from normal human breast tissue were isolated by immunomagnetic separation methods. Pools of RNA from highly enriched preparations of these cell types were subjected to expression profiling using massively parallel signature sequencing (MPSS) and four different genome wide microarray platforms. Functional related transcripts of the differential tumour epithelial transcriptome were used for gene set enrichment analysis to identify enrichment of luminal and myoepithelial type genes. Clinical pathological validation of a small number of genes was performed on tissue microarrays. MPSS identified 6,553 differentially expressed genes between the pool of normal luminal cells and that of primary tumours substantially enriched for epithelial cells, of which 98% were represented and 60% were confirmed by microarray profiling. Significant expression level changes between these two samples detected only by microarray technology were shown by 4,149 transcripts, resulting in a combined differential tumour epithelial transcriptome of 8,051 genes. Microarray gene signatures identified a comprehensive list of 907 and 955 transcripts whose expression differed between luminal epithelial cells and myoepithelial cells, respectively. Functional annotation and gene set enrichment analysis highlighted a group of genes related to skeletal development that were associated with the myoepithelial/basal cells and upregulated in the tumour sample. One of the most highly overexpressed genes in this category, that encoding periostin, was analysed immunohistochemically on breast cancer tissue microarrays and its expression in neoplastic cells correlated with poor outcome in a cohort of poor prognosis estrogen receptor-positive tumours. Using highly enriched cell populations in combination with multiplatform gene expression profiling studies, a comprehensive analysis of molecular changes between the normal and malignant breast tissue was established. This study provides a basis for the identification of novel and potentially important targets for diagnosis, prognosis and therapy in breast cancer.
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影响因子：
14.9
作者：
Draghici, S;Khatri, P;Tainsky, MA
通讯作者：
Tainsky, MA
影响因子：
2.9
作者：
OHARE, MJ;ORMEROD, MG;GUSTERSON, BA
通讯作者：
GUSTERSON, BA
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
11.2
作者：
Dai, HY;van't Veer, L;Friend, S
通讯作者：
Friend, S
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y