Mechanism of action of probucol on cholesteryl ester transfer protein (CETP) mRNA in a Chinese hamster ovary cell line that had been stably transfected with a human CETP gene

Jiafu Ou, Keijiro Saku, Shiro Jimi, Takao Ohta, Bo Zhang, Henry J. Pownall, Yoko Shimada, Yoshio Tsujita, Kikuo Arakawa

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Probucol, a widely used lipid-lowering agent, is associated with a significant reduction of plasma high density lipoprotein (HDL)-cholesterol levels. To examine the mechanism of probucol HDL-lowering and probucol's effects on cholesteryl ester transfer protein (CETP) and cholesterol metabolism in cells, we used a Chinese hamster ovary (CHO) cell line that had been stably transfected with a human CETP gene (hCETP-CHO). After this cell line was incubated with various concentrations of probucol (5, 10 and 50 μM) for 24 h, mean intracellular probucol concentrations reached 0.47, 0.67, and 1.52 μg/mg cell protein, respectively. Northern blot analysis showed that cellular CETP mRNA was increased by probucol in a dose-dependent manner (137%, 162%, and 221% of the control, respectively). The specific CET activity in the culture medium, measured as the percentage of [3H]cholesterol oleate transferred from discoidal bilayer particles (which mimic HDL) to LDL, also increased in a dose-dependent manner. Intracellular total cholesterol levels were decreased to 87.5%, 74.9%, and 52.5% of the control, respectively. Probucol had no effects on HMG-CoA reductase activity or cholesterol synthesis from [14C]acetate in hCETP-CHO. However, 14C-incorporated cholesterol secretion into the culture medium from hCETP-CHO was increased to 181%, 256% and 354% of the control by 5, 10 and 50 μM probucol, respectively. We concluded that (1) treatment with probucol increased the CETP mRNA level and specific CET activity in the hCETP-CHO cell line, and (2) probucol promoted cholesterol efflux from hCETP-CHO, which resulted in a decrease in intracellular cholesterol levels. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)153-160
Number of pages8
JournalBiochimica et Biophysica Acta - Lipids and Lipid Metabolism
Volume1393
Issue number1
DOIs
StatePublished - Jul 31 1998

Keywords

  • Chinese hamster ovary cell line
  • Cholesteryl ester transfer protein
  • Lipid
  • Lipoprotein
  • mRNA
  • Probucol

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Biophysics

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