Inhibition of human-type 1 3beta-hydroxysteroid deshydrogenase/Delta(5)-Delta(4)-isomerase expression using siRNA.
Specific inhibition of type 1 3beta-HSD is of particular interest since it will allow us to control the formation of androgens and estrogens in peripheral target tissues without affecting type 2 3beta-HSD, which is responsible for the biosynthesis of glucocorticoids and mineralocorticoids in the adrenals. The high homology between types 1 and 2 3beta-HSD is a major difficulty in the development of specific inhibitors through classical chemical synthesis. In this report, we describe the use of small interference RNA (siRNA) to specifically inhibit human type 1 3beta-HSD. We have constructed three DNA vector-based RNAi vectors that allow us to produce three RNA duplexes of 21 nucleotides targeting three different coding regions of human type 1 3beta-HSD mRNA. The resulting constructs were co-transfected into HEK-293 cells with a vector expressing type 1 3beta-HSD. The results indicate that while the two duplexes that target sequences in the 5'-region do not have a strong inhibitory effect, the duplex that targets the 3'-region efficiently inhibits 3beta-HSD activity. Up to 98% inhibition has been observed. To our knowledge, this is the first report showing successful inhibition of steroidogenic enzymes using siRNA technology.
Samson
Mélanie
M
Oncology and Molecular Endocrinology Research Center, Laval University Medical Center, 2705 Laurier Boulevard, Que., Canada G1V 4G2.
Labrie
Fernand
F
Luu-The
Van
V
D015096:3-Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors* / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D007668:Kidney; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000494:pharmacology*; D015183:Restriction Mapping
D004791:Enzyme Inhibitors; D034741:RNA, Small Interfering; D015096:3-Hydroxysteroid Dehydrogenases