Altered morphology in cultured rat intestinal epithelial IEC-6 cells is associated with alkaline phosphatase expression |
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Authors: | Wood S R Zhao Q Smith L H Daniels C K |
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Affiliation: | Department of Pharmaceutical Sciences, College of Pharmacy, Idaho State University, Box 8334, Pocatello, ID 83209, USA. |
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Abstract: | Non-transformed, rat intestinal epithelial cells (IEC-6), and human intestinal colonic carcinoma cells (CACO-2) have both been used to study processes of epithelial cell differentiation. However, only CACO-2 cells have been described as spontaneously expressing phenotypic changes of differentiation in culture. We report here that when IEC-6 cells are grown in post-confluent culture, they develop structural changes similar to those seen in cells induced to differentiate by culture on Englebreth-Holm-Swarm (EHS) extracellular matrix proteins. Correlated with this morphological change is loss of nuclear localization of c-myc protein and development of cell surface alkaline phosphatase (ALP) enzymatic activity. Messenger RNAs for liver and intestinal isoforms of ALP were expressed in both pre- and post-confluent cells. Inhibition of ALP activity in post-confluent cells by levamisole indicated the expressed ALP activity to be of the liver isoform. We suggest the expression of ALP activity, which occurs concomitantly with morphological alterations in post-confluent IEC-6 cells, represents increased expression and localization to the cell surface of the liver isoform of ALP. Cultured IEC-6 cells may provide a non-transformed, in vitro alternative to CACO-2 cells for study of epithelial cell differentiation. |
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Keywords: | Intestinal epithelial cell Alkaline phosphatase Immunohistochemistry Gene expression IEC-6 cells |
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