Analysis of gene expression in prostate cancer epithelial and interstitial stromal cells using laser capture microdissection.
Analysis of gene expression in prostate cancer epithelial and interstitial stromal cells using laser capture microdissection.
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DOI：
10.1186/1471-2407-10-165
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发表时间：
2010-04-28
期刊：
影响因子：
3.8
通讯作者：
Fraizer GC
中科院分区：
文献类型：
--
作者：
Gregg JL;Brown KE;Mintz EM;Piontkivska H;Fraizer GC
前列腺是一个多方面的系统，在这个系统中，前列腺上皮和间质具有不同的生理作用。为了了解间质和腺上皮之间的相互作用，有必要描绘这两种组织类型在前列腺癌中的基因表达谱。大多数研究通过使用混合细胞群进行全局表达分析来比较肿瘤和正常样本。这份报告提出了第一项关于前列腺癌组织的研究，该研究使用激光捕获显微解剖(LCM)检查特定细胞类型之间的差异表达模式。LCM被用来分离不同的细胞类型群体，并使用寡核苷酸微阵列鉴定它们的基因表达差异。然后用实时定量聚合酶链式反应分析配对的前列腺癌和非肿瘤性前列腺组织中的10个差异表达基因。进一步比较了转录因子WT1和Egr1在已建立的前列腺细胞系中的表达模式。用免疫组织化学方法检测前列腺组织芯片中WT1蛋白的表达。激光捕获和微阵列分析的两步法确定了近500个基因，它们在前列腺上皮组织和间质组织中的表达水平存在显著差异。在上皮细胞中表达的几个基因(WT1、GATA2和FGFR-3)在肿瘤组织中的表达高于非肿瘤组织；相反，在间质细胞中表达的几个基因(CCL5、CXCL13、IGF-1、FGF-2和IGFBP3)在非肿瘤组织中的表达高于肿瘤组织。值得注意的是，Egr1在上皮组织和间质组织中的表达也存在差异。WT1和Egr1在细胞系中的表达与这些差异表达模式一致。重要的是，WT1蛋白在肿瘤组织中表达，而在正常组织和良性组织中不表达。前列腺代表了多种细胞类型的复杂混合，有必要分析不同的细胞群，以更好地了解它们之间的潜在相互作用。在本研究中，我们使用激光共聚焦显微镜和微阵列分析来鉴定前列腺细胞群体中新的基因表达模式，包括鉴定上皮细胞中WT1的表达。通过对肿瘤组织和细胞系的分析，证实了WT1在前列腺癌中的表达相关性，提示WT1在前列腺癌的发生中具有潜在的作用。
The prostate gland represents a multifaceted system in which prostate epithelia and stroma have distinct physiological roles. To understand the interaction between stroma and glandular epithelia, it is essential to delineate the gene expression profiles of these two tissue types in prostate cancer. Most studies have compared tumor and normal samples by performing global expression analysis using a mixture of cell populations. This report presents the first study of prostate tumor tissue that examines patterns of differential expression between specific cell types using laser capture microdissection (LCM). LCM was used to isolate distinct cell-type populations and identify their gene expression differences using oligonucleotide microarrays. Ten differentially expressed genes were then analyzed in paired tumor and non-neoplastic prostate tissues by quantitative real-time PCR. Expression patterns of the transcription factors, WT1 and EGR1, were further compared in established prostate cell lines. WT1 protein expression was also examined in prostate tissue microarrays using immunohistochemistry. The two-step method of laser capture and microarray analysis identified nearly 500 genes whose expression levels were significantly different in prostate epithelial versus stromal tissues. Several genes expressed in epithelial cells (WT1, GATA2, and FGFR-3) were more highly expressed in neoplastic than in non-neoplastic tissues; conversely several genes expressed in stromal cells (CCL5, CXCL13, IGF-1, FGF-2, and IGFBP3) were more highly expressed in non-neoplastic than in neoplastic tissues. Notably, EGR1 was also differentially expressed between epithelial and stromal tissues. Expression of WT1 and EGR1 in cell lines was consistent with these patterns of differential expression. Importantly, WT1 protein expression was demonstrated in tumor tissues and was absent in normal and benign tissues. The prostate represents a complex mix of cell types and there is a need to analyze distinct cell populations to better understand their potential interactions. In the present study, LCM and microarray analysis were used to identify novel gene expression patterns in prostate cell populations, including identification of WT1 expression in epithelial cells. The relevance of WT1 expression in prostate cancer was confirmed by analysis of tumor tissue and cell lines, suggesting a potential role for WT1 in prostate tumorigenesis.
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