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Journal of Lipid Research, Vol 28, 1350-1363, Copyright © 1987 by Lipid Research, Inc.
MG Traber, HJ Kayden and MJ Rindler
Lipoprotein secretion by Caco-2 cells, a human intestinal cell line, was
studied in cells grown on inserts containing a Millipore filter (0.45
micron), separating secretory products from the apical and basolateral
membranes into separate chambers. Under these conditions, as observed by
electron microscopy, the cells formed a monolayer of columnar epithelial
cells with microvilli on the apical surface and tight junctions between
cells. The electrical resistances of the cell monolayers were 250-500
ohms/cm2. Both 14C-labeled lipids and 35S- labeled proteins were used to
assess lipoprotein secretion. After a 24- hr incubation with [14C]oleic
acid, 60-80% of the secreted triglyceride (TG) was in the basolateral
chamber; 40% of the TG was present in the d less than 1.006 g/ml
(chylomicron + VLDL) fraction and 50% in the 1.006 less than d less than
1.063 g/ml (LDL) fraction. After a 4-hr incubation with [35S]methionine,
apolipoproteins were found to be major secretory products with 75-100%
secreted to the basolateral chamber. Apolipoproteins B-100, B-48, E, A-I,
A-IV, and C-III were identified by immunoprecipitation. The d less than
1.006 g/ml fraction was found to contain all of the major apolipoproteins,
while the LDL fraction contained primarily apoB-100 and apoE; the HDL
(1.063 less than d less than 1.21 g/ml) fraction principally contained
apoA-I and apoA-IV. Mn- heparin precipitated all of the
[35S]methionine-labeled apoB-100 and B- 48 and a majority of the other
apolipoproteins, and 80% of the [14C]oleic acid-labeled triglyceride, but
only 15% of the phospholipid, demonstrating that Caco-2 cells secrete
triglyceride-rich lipoproteins containing apoB. Secretion of lipoproteins
was dependent on the lipid content of the medium; prior incubation with
lipoprotein-depleted serum specifically reduced the secretion of
lipoproteins, while addition of both LDL and oleic acid to the medium
maintained the level of apoB-100, B-48, and A-IV secretion to that observed
in the control cultures.
ARTICLES
Polarized secretion of newly synthesized lipoproteins by the Caco-2 human intestinal cell line
Department of Medicine, New York University School of Medicine, NY 10016.
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