4.6 Article

Quantitative analysis of the expression of ACAT1 genes in human tissues by real-time PCR2

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JOURNAL OF LIPID RESEARCH
卷 45, 期 4, 页码 686-696

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ELSEVIER
DOI: 10.1194/jlr.M300365-JLR200

关键词

acyl-coenzyme A : cholesterol acyltransferase; atherosclerosis; cholesterol metabolism; duodenum; gallstones; intestine; liver; sterol o-acyltransferase

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ACAT (also called sterol o-acyltransferase) catalyzes the esterification of cholesterol by reaction with longchain acyl-CoA derivatives and plays a pivotal role in the regulation of cholesterol homeostasis. Although two human ACAT genes termed ACAT-1 and ACAT-2 have been reported, prior research on differential tissue expression is qualitative and incomplete. We have developed a quantitative multiplex assay for each ACAT isoform after RT treatment of total RNA using TaqMan real-time quantitative PCR normalized to beta-actin in the same reaction tube. This enabled us to calculate the relative abundance of transcripts in several human tissues as an ACAT 2/ACAT-1 ratio. In liver (n = 17), ACAT-1 transcripts were on average 9-fold (range, 1.7- to 167-fold) more abundant than ACAT-2, whereas in duodenal samples (n = 10), ACAT-2 transcripts were on average 3-fold (range, 0.39- to 12.2-fold) more abundant than ACAT-1. ACAT-2 was detected for the first time in peripheral blood mononuclear cells. Interesting differences in ACAT-2 mRNA expression were evident in subgroup analysis of samples from different sources. These results demonstrate quantitatively that ACAT-1 transcripts predominate in human liver and ACAT-2 transcripts predominate in human duodenum and support the notion that ACAT-2 has an important regulatory role in liver and intestine.

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