Quantitative analysis of the expression of ACAT 1 genes in human tissues by real-time PCR2

Journal of Lipid Research

卷:45

Robert Simpson¹  Shaun P. Scott¹  Kavitha Rangaraj²  Katherine A. Stuart²  Grant A. Ramm³  Donald J. Maclean³  Les K. Nathanson⁴  Jeffery L. Smith⁴  John de Jersey⁴ 

[1] Department of Gastroenterology and Hepatology, Princess Alexandra Hospital, Brisbane 4102, Australia;

[2] Department of Surgery, University of Queensland, Brisbane 4072, Australia;

[3] The Queensland Institute of Medical Research, Brisbane 4006, Australia;

[4] Department of Biochemistry and Molecular Biology, University of Queensland, Brisbane 4072, Australia;

关键词: acyl-coenzyme A:cholesterol acyltransferase;    atherosclerosis;    cholesterol metabolism;    duodenum;    gallstones;    intestine;   

DOI  :  

来源: DOAJ

【 摘 要 】

ACAT (also called sterol o-acyltransferase) catalyzes the esterification of cholesterol by reaction with long-chain 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 β-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.

【 授权许可】

Unknown