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Updated: Aug 12, 2026

Cholesterol Efflux Assay
Published on: March 6, 2012
Effect of 25-hydroxycholesterol on cholesteryl ester formation in Caco-2 cells
H Kusuhara1, O Shimada, J Inui
1Research Laboratories, Yoshitomi Pharmaceutical Industries, Ltd., Saitama, Japan.
Abstract:
Incubation of Caco-2 cells, a human intestinal cell line, with 25-hydroxycholesterol (25-HOC) marked enhanced cellular cholesteryl ester formation determined by incorporation of [14C]oleic acid into intracellular cholesteryl [14C]oleate. The stimulation by 25-HOC of cholesteryl ester formation was suppressed by staurosporine, a kinase inhibitor, but not by cycloheximide or actinomycin D. The specific activity of microsomal acyl-coenzyme A:cholesterol acyltransferase (ACAT) increased two-fold in cells treated with 10 microM 25-HOC for 5 h. ACAT activity decreased when microsomes were incubated without sodium fluoride, a phosphatase inhibitor, but the decrease in ACAT activity in cells stimulated with 25-HOC was more pronounced. The results suggest that protein phosphorylation may be involved in the stimulation of cholesteryl ester formation by 25-HOC in Caco-2 cells.
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