Landowski Christopher P, Anderle Pascale, Sun Duxin, Sadee Wolfgang, Amidon Gordon L
Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA.
AAPS J. 2004 Sep 7;6(3):e21. doi: 10.1208/aapsj060321.
mRNA expression profiles had previously been measured in Caco-2 cells (human colonic carcinoma cells) using either custom-designed spotted oligonucleotide arrays or Affymetrix GeneChip oligonucleotide arrays. The Caco-2 cells used were from different clones and were examined under slightly different culture conditions commonly encountered when Caco-2 cells are used as a model tissue for studying intestinal transport and metabolism in different laboratories. In this study, we compared gene expression profiles of Caco-2 cells generated with different arrays to assess the validity of conclusions derived from the 2 independent studies, with a focus on changes in transporter and ion channel mRNA expression levels on Caco-2 cell differentiation. Significant changes in expression levels upon differentiation were observed with 78 genes, with probes common to both arrays. Of these, 18 genes were upregulated and 36 genes were downregulated. The 2 arrays yielded discrepant results for 24 genes, showing significant changes upon differentiation. The results from the 2 arrays correlated well for genes expressed above average levels (r = 0.75, P < 0.01, n = 25) and poorly for genes expressed at low levels (r = 0.08, P > 0.05, n = 25). Overall correlation across the 2 platforms was r = 0.45 (P < 0.01) for the 78 genes, with similar results from both arrays. Despite differences in experimental conditions and array technology, similar results were obtained for most genes.
此前,使用定制设计的点阵寡核苷酸阵列或Affymetrix基因芯片寡核苷酸阵列,在Caco-2细胞(人结肠癌细胞)中测量了mRNA表达谱。所使用的Caco-2细胞来自不同克隆,并且是在不同实验室将Caco-2细胞用作研究肠道转运和代谢的模型组织时通常遇到的略有不同的培养条件下进行检测的。在本研究中,我们比较了用不同阵列生成的Caco-2细胞的基因表达谱,以评估来自两项独立研究的结论的有效性,重点关注Caco-2细胞分化时转运体和离子通道mRNA表达水平的变化。在78个基因中观察到分化时表达水平的显著变化,这两个阵列有共同的探针。其中,18个基因上调,36个基因下调。对于24个基因,这两个阵列产生了不一致的结果,显示出分化时的显著变化。对于表达水平高于平均水平的基因,两个阵列的结果相关性良好(r = 0.75,P < 0.01,n = 25),而对于低水平表达的基因,相关性较差(r = 0.08,P > 0.05,n = 25)。对于这78个基因,两个平台之间的总体相关性为r = 0.45(P < 0.01),两个阵列的结果相似。尽管实验条件和阵列技术存在差异,但大多数基因获得了相似的结果。