Tremblay Eric, Auclair Joëlle, Delvin Edgar, Levy Emile, Ménard Daniel, Pshezhetsky Alexey V, Rivard Nathalie, Seidman Ernest G, Sinnett Daniel, Vachon Pierre H, Beaulieu Jean-François
CIHR Group on Functional Development and Physiopathology of the Digestive Tract, Sherbrooke, Québec, Canada.
J Cell Biochem. 2006 Nov 1;99(4):1175-86. doi: 10.1002/jcb.21015.
cDNA microarray technology enables detailed analysis of gene expression throughout complex processes such as differentiation. The aim of this study was to analyze the gene expression profile of normal human intestinal epithelial cells using cell models that recapitulate the crypt-villus axis of intestinal differentiation in comparison with the widely used Caco-2 cell model. cDNA microarrays (19,200 human genes) and a clustering algorithm were used to identify patterns of gene expression in the crypt-like proliferative HIEC and tsFHI cells, and villus epithelial cells as well as Caco-2/15 cells at two distinct stages of differentiation. Unsupervised hierarchical clustering analysis of global gene expression among the cell lines identified two branches: one for the HIEC cells versus a second comprised of two sub-groups: (a) the proliferative Caco-2 cells and (b) the differentiated Caco-2 cells and closely related villus epithelial cells. At the gene level, supervised hierarchical clustering with 272 differentially expressed genes revealed distinct expression patterns specific to each cell phenotype. We identified several upregulated genes that could lead to the identification of new regulatory pathways involved in cell differentiation and carcinogenesis. The combined use of microarray analysis and human intestinal cell models thus provides a powerful tool for establishing detailed gene expression profiles of proliferative to terminally differentiated intestinal cells. Furthermore, the molecular differences between the normal human intestinal cell models and Caco-2 cells clearly point out the strengths and limitations of this widely used experimental model for studying intestinal cell proliferation and differentiation.
cDNA微阵列技术能够对诸如分化等复杂过程中的基因表达进行详细分析。本研究的目的是使用能够重现肠道分化隐窝-绒毛轴的细胞模型，与广泛使用的Caco-2细胞模型相比较，分析正常人肠道上皮细胞的基因表达谱。利用cDNA微阵列（19,200个人类基因）和聚类算法，来识别隐窝样增殖性人小肠上皮细胞（HIEC）和tsFHI细胞、绒毛上皮细胞以及处于两个不同分化阶段的Caco-2/15细胞中的基因表达模式。对各细胞系间的整体基因表达进行无监督层次聚类分析，识别出两个分支：一个是HIEC细胞分支，另一个分支由两个亚组组成：(a) 增殖性Caco-2细胞，以及(b) 分化的Caco-2细胞和密切相关的绒毛上皮细胞。在基因水平上，对272个差异表达基因进行有监督层次聚类，揭示了每种细胞表型特有的不同表达模式。我们鉴定出了几个上调基因，这些基因可能有助于识别参与细胞分化和癌变的新调控途径。因此，微阵列分析与人类肠道细胞模型的联合应用为建立从增殖到终末分化的肠道细胞的详细基因表达谱提供了一个强大工具。此外，正常人肠道细胞模型与Caco-2细胞之间的分子差异清楚地指出了这个广泛用于研究肠道细胞增殖和分化的实验模型的优势和局限性。
