Net transport of cholesterol from cells of the human EA.hy 926 endothelial cell line to high density lipoproteins.

EA.hy 926 cells, a human endothelial cell line, show characteristics of differentiated endothelial cells. The cells express saturable binding of apo E-free 125I-high density lipoprotein3 (HDL3). Bmax increased from 71 to 226 ng HDL3 bound/mg cell protein after cholesterol loading of the confluent endothelial cells with cationized low density lipoprotein (LDL). The affinity did not change after cholesterol enrichment (Kd was 37 micrograms HDL3 protein/ml for control cells and 31 micrograms/ml for loaded cells). Incubation of cholesterol-loaded EA.hy 926 cells with native HDL and LDL had different effects on cellular cholesterol levels. Incubation with HDL decreased both esterified and unesterified cellular cholesterol, but LDL did not change total cellular cholesterol. However, LDL tended to increase cellular cholesteryl esters, with a concomitant decrease of unesterified cellular cholesterol. Incubation of endothelial cells with both HDL and LDL also resulted in decreased total cellular cholesterol levels. These data show that cationized LDL-loaded human endothelial EA.hy 926 cells can be used to study the net transport of cellular cholesterol to HDL, the first step in reverse cholesterol transport.