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The human intestinal cell lines Caco-2 and LS174T as models to study cell-type specific mucin expression
Authors:B. Jan-Willem van Klinken  Esmeralda Oussoren  Jan-Johan Weenink  Ger J. Strous  Hans A. Büller  Jan Dekker  Alexandra W. C. Einerhand
Affiliation:(1) Pediatric Gastroenterology and Nutrition, Academic Medical Center, Room G8-260, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands;(2) Department of Cell Biology, Medical School, Utrecht, The Netherlands
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 (sim550 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 (sim470 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.Abbreviations SDS-PAGE sodium dodecylsulfate-polyacrylamide gel electrophoresis - DMEM Dulbecco's modified Eagle's medium - EMEM Eagle's minimum essential medium - Endo-H endo-beta-N-acetylglucosaminidase H - HGBM human gallbladder mucin - dpc days past confluence - PBS phosphate buffered saline
Keywords:mucin-type glycoproteins  MUC1-6 mRNA biosynthesis  protein biosynthesis  Caco-2  LS174T  human intestine
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