Transcriptional regulation of endothelial constitutive PGHS-1 expression by phorbol ester

• Published in: The American journal of physiology Vol. 270; no. 1 Pt 1; p. C259
• Main Authors: Xu, X M, Tang, J L, Hajibeigi, A, Loose-Mitchell, D S, Wu, K K
• Format: Journal Article
• Language: English
• Published: United States 01.01.1996
• Subjects: Base Sequence; Blotting, Northern; Cells, Cultured; Endothelium, Vascular - cytology; Endothelium, Vascular - metabolism; Humans; Molecular Probes - genetics; Molecular Sequence Data; Osmolar Concentration; Polymerase Chain Reaction; Promoter Regions, Genetic; Prostaglandin-Endoperoxide Synthases - genetics; Prostaglandin-Endoperoxide Synthases - metabolism; RNA, Messenger - metabolism; Tetradecanoylphorbol Acetate - pharmacology; Time Factors; Transcription, Genetic - drug effects
• ISSN: 0002-9513
• DOI: 10.1152/ajpcell.1996.270.1.C259

Human endothelial cells contain two isoforms of prostaglandin H synthase (PGHS). PGHS-1 is constitutively expressed, whereas PGHS-2 is inducible. To determine whether expression of PGHS-1 is regulated, we treated cultured human umbilical vein endothelial cells (HUVEC) with phorbol 12-myristate 13-acetate (PMA) or its inactive analogue and measured PGHS-1 mRNA levels by Northern analysis and competitive polymerase chain reaction. PMA increased PGHS-1 mRNA levels determined by both techniques in a time- and concentration-dependent manner. The mRNA level was increased about twofold over the basal level after 4-6 h of PMA (10-50 nM) treatment. The level of PGHS-1 protein was similarly increased by PMA. Stimulation of PGHS-1 mRNA levels was abrogated by cycloheximide, actinomycin D, staurosporine, or calphostin C. The 5'-promoter activity of human PGHS-1 gene was increased twofold over the basal level by PMA in NS-20 cells. These results indicate that the constitutive PGHS-1 in HUVEC is transcriptionally stimulated by PMA in a protein kinase C-dependent manner.