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J Lipid Res. 2018 Oct;59(10):1940-1950. doi: 10.1194/jlr.M087635. Epub 2018 Aug 16.

Phosphatidylcholine synthesis regulates triglyceride storage and chylomicron secretion by Caco2 cells.

Author information

1
Atlantic Research Center, Departments of Pediatrics, and Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
2
Atlantic Research Center, Departments of Pediatrics, and Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada nridgway@dal.ca.

Abstract

Intracellular lipid droplets (LDs) supply fatty acids for energy, membrane biogenesis, and lipoprotein secretion. The surface monolayer of LDs is composed of phospholipids, primarily phosphatidylcholine (PC), that stabilize the neutral lipid core of triglyceride (TG). To determine the relationship between PC synthesis and TG storage and secretion in chylomicrons, we used a model of intestinal-derived human epithelial colorectal adenocarcinoma (Caco2) cells with knockout of PCYT1A, which encodes the rate-limiting enzyme CTP:phosphocholine cytidylyltransferase (CCT)α in the CDP-choline pathway, that were treated with the fatty acid oleate. CRISPR/Cas9 knockout of CCTα in Caco2 cells (Caco2-KO cells) reduced PC synthesis by 50%. Compared with Caco2 cells, Caco2-KO cells exposed to oleate had fewer and larger LDs and greater TG accumulation as a result. The addition of exogenous lysophosphatidylcholine to Caco2-KO cells reversed the LD morphology defect. Caco2-KO cells, differentiated into epithelial monolayers, accumulated intracellular TG and had deficient TG and chylomicron-associated apoB48 secretion; apoB100 secretion was unaffected by CCTα knockout or oleate. Metabolic-labeling and LD imaging of Caco2-KO cells indicated preferential shuttling of de novo synthesized TG into larger LDs rather than into chylomicrons. Thus, reduced de novo PC synthesis in Caco2 cells enhances TG storage in large LDs and inhibits apoB48 chylomicron secretion.

KEYWORDS:

cytidine triphosphate:phosphocholine cytidylyltransferase; lipid droplets; lipoproteins/assembly; phospholipids; triglycerides

PMID:
30115754
PMCID:
PMC6168309
DOI:
10.1194/jlr.M087635
[Indexed for MEDLINE]
Free PMC Article

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