Lipid, apolipoprotein, and lipoprotein synthesis and secretion during cellular differentiation in Caco-2 cells.
Although Caco-2 cells are frequently employed for the study of enterocyte lipid metabolism, variable results have been reported regarding their ability to synthesize and secrete lipids and apolipoproteins. The major goal of this investigation is to examine the capacity of Caco-2 cells to elaborate and secrete lipids, lipoproteins, and apolipoproteins at different degrees of morphological and functional differentiation. Cells were cultured in medium with 5% fetal bovine serum (FBS), on permeable polycarbonate filters from 2 to 30 d in the presence of 14C-oleate or 35S-methionine. Cellular differentiation, as assessed by morphology (light and electron microscopy), transepithelial resistance, free fatty acid flux, and sucrase activity, progressed steadily up to 20 d of culture. Caco-2 cells esterified oleic acid mainly into phospholipids, triglycerides (TG), and smaller amounts of cholesterol esters. Lipid synthesis began as early as 2 d, and TG secretion was enhanced with increased duration of culture. However, very low efficiency of lipid export was observed at all levels of differentiation, reaching a maximum of only 6% of intracellular lipids. VLDL and LDL were the dominant lipoproteins secreted, with HDL comprising < 20% of the total. VLDL secretion increased, while LDL decreased, whereas the lipid composition of lipoproteins varied little with increasing duration of culture. Apoprotein B and A-I synthesis and secretion increased markedly from 11 to 20 d of culture. The ratio of apo B-100/B-48 decreased between 11 and 30 d, consistent with enhanced apo B editing of more mature enterocytes. Taken together, our data suggest that from 20 d of culture, Caco-2 cells are morphologically and functionally mature, capable of lipid esterification, and lipoprotein and apolipoprotein synthesis. However, despite their functional and morphological similarities to mature enterocytes, Caco-2 cells have a very limited lipid export capacity.
Mehran
M
M
Hôpital Ste.-Justine, Department of Pediatrics, Faculty of Medicine, Université de Montréal, Quebec, Canada.
Levy
E
E
Bendayan
M
M
Seidman
E
E
D001053:Apolipoproteins / Q000096:biosynthesis* / Q000378:metabolism; D018938:Caco-2 Cells / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D002454:Cell Differentiation; D006801:Humans; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis / Q000096:biosynthesis*; D008074:Lipoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D019301:Oleic Acid / Q000378:metabolism
D001053:Apolipoproteins; D008055:Lipids; D008074:Lipoproteins; D019301:Oleic Acid