Messenger RNA expression of transporter and ion channel genes in undifferentiated and differentiated Caco-2 cells compared to human intestines
Messenger RNA expression of transporter and ion channel genes in undifferentiated and differentiated Caco-2 cells compared to human intestines
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DOI：
10.1023/a:1022282221530
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发表时间：
2003-01-01
影响因子：
3.7
通讯作者：
Sadée, W
中科院分区：
文献类型：
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作者：
Anderle, P;Rakhmanova, V;Sadée, W
目的。本工作的目的是研究细胞分化对 Caco-2 细胞中转运蛋白和通道 mRNA 表达的影响，并评估 Caco-2 细胞作为载体介导的肠道药物转运模型的方法。使用带有 750 个脱氧寡核苷酸探针（70mers）的定制设计微阵列芯片测量基因 mRNA 表达。每种低聚物在聚赖氨酸包被的载玻片上印刷四次。表达谱表示为源自 Caco-2 细胞的 Poly(A) + RNA 样品的 Cy3 和 Cy5 染料标记的 cDNA 与人肠总 RNA 的荧光强度之间的比率值。结果。 Caco-2 细胞在培养 5 天至 2 周的分化过程中观察到转运蛋白和通道 mRNA 表达谱的显着差异，包括 MAT8、S 蛋白和 Nramp2 的变化。比较不同传代数的Caco-2细胞发现，除了GLUT3之外，mRNA几乎没有变化，GLUT3在13个传代数内下调了2.4倍。无论 Caco-2 细胞的分化状态如何，Caco-2 细胞在烧瓶中或过滤器上培养时都具有相似的表达谱，但与人小肠和大肠的差异更大。在小肠或大肠中高度转录的几种基因的表达在 Caco-2 细胞中相差四倍或更多。结论。尽管 Caco-2 细胞已被证明是研究人类肠道中载体介导的转运的合适模型，但特定转运蛋白和离子通道基因的表达可能存在很大差异。
Purpose. The purpose of this work was to study the influence of cell differentiation on the mRNA expression of transporters and channels in Caco-2 cells and to assess Caco-2 cells as a model for carrier-mediated drug transport in the intestines.Method. Gene mRNA expression was measured using a custom-designed microarray chip with 750 deoxyoligonucleotide probes (70mers). Each oligomer was printed four times on poly-lysine-coated glass slides. Expression profiles were expressed as ratio values between fluorescence intensities of Cy3 and Cy5 dye-labeled cDNA derived from poly(A) + RNA samples of Caco-2 cells and total RNA of human intestines.Results. Significant differences in the mRNA expression profile of transporters and channels were observed upon differentiation of Caco-2 cells from 5 days to 2 weeks in culture, including changes for MAT8, S-protein, and Nramp2. Comparing Caco-2 cells of different passage number revealed few changes in mRNAs except for GLUT3, which was down-regulated 2.4-fold within 13 passage numbers. Caco-2 cells had a similar expression profile when either cultured in flasks or on filters but differed more strongly from human small and large intestine, regardless of the differentiation state of Caco-2 cells. Expression of several genes highly transcribed in small or large intestines differed fourfold or more in Caco-2 cells.Conclusions. Although Caco-2 cells have proven a suitable model for studying carrier-mediated transport in human intestines, the expression of specific transporter and ion channel genes may differ substantially.