The regulation of apoB synthesis and secretion by lipids present within bile was investigated in CaCo-2 cells grown on semipermeable filters. Bile acids decreased the basolateral secretion of immunoreactive apoB. Taurocholic acid decreased the secretion of newly synthesized apoB by increasing the rate of apoB degradation, but had no effect on the synthesis and secretion of apoA-I or trichloroacetic acid-precipitable proteins. The calcium ionophore, A23187, decreased apoB secretion similar to that observed for taurocholate. The addition of the ionophore and taurocholate together did not cause a further decrease in apoB secretion. Cholesterol or its hydroxylated derivative, 25-hydroxycholesterol, did not alter secretion of immunoreactive or newly synthesized apoB. Phosphatidylcholine increased apoB synthesis and secretion without affecting the synthesis or secretion of apoA-I. Phosphatidylcholine also reversed the effect of A23187 on apoB secretion. When phosphatidylcholine was added to the basolateral medium, apoB secretion was not altered. ApoB secretion was not increased by phospholipids of other classes. Dioleoylphosphatidylcholine increased apoB secretion, whereas dipalmitoylphosphatidylcholine did not. Fatty acid-labeled phosphatidylcholine was not hydrolyzed in the apical medium. Only 2% of the added phosphatidylcholine was cell-associated, and of this, 80% of the label remained as phosphatidylcholine with most of the remainder in triacylglycerols, fatty acids, and phosphatidylethanolamine. The results suggest that bile acids decrease apoB secretion by increasing its rate of degradation. This effect may be related to their ionophoric property. Cholesterol flux does not regulate apoB secretion. Phosphatidylcholine, independent of triacylglycerol flux and independent of its hydrolysis, increases the secretion of apoB by increasing apoB synthesis. Luminal phosphatidylcholine may play a role in apoB secretion in the intestine.