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Messenger RNA Expression of Transporter and Ion Channel Genes in Undifferentiated and Differentiated Caco-2 Cells Compared to Human Intestines

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Abstract

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.

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Authors and Affiliations

  1. Department of Biopharmaceutical Sciences, School of Pharmacy, University of San Francisco, San Francisco, California

    Pascale Anderle, Vera Rakhmanova & Wolfgang Sadée

  2. Xenoport Inc, Palo Alto, California

    Katie Woodford & Noa Zerangue

  3. Department of Pharmacology, College of Medicine and Public Health, Ohio State University, Columbus, Ohio

    Pascale Anderle & Wolfgang Sadée

  4. ISREC, Epalinges s/Lausanne, Switzerland

    Pascale Anderle

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  1. Pascale Anderle
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  2. Vera Rakhmanova
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Correspondence to Pascale Anderle.

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Anderle, P., Rakhmanova, V., Woodford, K. et al. Messenger RNA Expression of Transporter and Ion Channel Genes in Undifferentiated and Differentiated Caco-2 Cells Compared to Human Intestines. Pharm Res 20, 3–15 (2003). https://doi.org/10.1023/A:1022282221530

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