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The human intestinal cell lines Caco-2 and LS174T as models to study cell-type specific mucin expression

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Abstract

Mucin expression was studied during proliferation and differentiation of the enterocyte-like Caco-2 and goblet cell-like LS174T cell lines. Caco-2 cells express mRNAs of MUC1, MUC3, MUC4 and MUC5A/C whereas MUC2 and MUC6 mRNAs are virtually absent. Furthermore, MUC3 mRNA is expressed in a differentiation dependent manner, as is the case for enterocytes. Concomitantly MUC3 protein precursor (∼550 kDa) was detected in Caco-2 cells. In LS174T cells mucin mRNAs of MUC1, MUC2 and MUC6 are constitutively expressed at high levels, whereas MUC3, MUC4 and MUC5A/C mRNAs are present at low levels. At the protein level LS174T cells express the goblet cell specific mucin protein precursors MUC2, MUC5A/C and MUC6 with apparent molecular masses of about 600 kDa, 470/500 kDa and 400 kDa respectively. MUC3 protein is not detectable. Furthermore, human gallbladder mucin protein (∼470 kDa precursor), of which the gene has not yet been identified, is expressed in LS174T cells. In addition, synthesis and secretion of the goblet cell specific mature MUC2, MUC5A/C and human gallbladder mucin was demonstrated in LS174T cells. It is concluded that Caco-2 and LS174T cell lines provide excellentin vitro models to elucidate the cell-type specific mechanisms responsible for mucin expression.

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Abbreviations

SDS-PAGE:

sodium dodecylsulfate-polyacrylamide gel electrophoresis

DMEM:

Dulbecco's modified Eagle's medium

EMEM:

Eagle's minimum essential medium

Endo-H:

endo-β-N-acetylglucosaminidase H

HGBM:

human gallbladder mucin

dpc:

days past confluence

PBS:

phosphate buffered saline

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Authors and Affiliations

  1. Pediatric Gastroenterology and Nutrition, Academic Medical Center, Room G8-260, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands

    B. Jan-Willem van Klinken, Esmeralda Oussoren, Jan-Johan Weenink, Hans A. Büller, Jan Dekker & Alexandra W. C. Einerhand

  2. Department of Cell Biology, Medical School, Utrecht, The Netherlands

    Ger J. Strous

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  1. B. Jan-Willem van Klinken
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  2. Esmeralda Oussoren
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  3. Jan-Johan Weenink
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  7. Alexandra W. C. Einerhand
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van Klinken, B.JW., Oussoren, E., Weenink, JJ. et al. The human intestinal cell lines Caco-2 and LS174T as models to study cell-type specific mucin expression. Glycoconjugate J 13, 757–768 (1996). https://doi.org/10.1007/BF00702340

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  • DOI: https://doi.org/10.1007/BF00702340

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