Inhibition of expression of protein kinase C alpha by antisense cDNA inhibits phorbol ester-mediated arachidonate release

• Published in: The Journal of biological chemistry Vol. 268; no. 16; pp. 11946 - 11950
• Main Authors: GODSON, C, BELL, K. S, INSEL, P. A
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 05.06.1993
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Arachidonic Acid - metabolism; Biological and medical sciences; Blotting, Northern; Blotting, Southern; Blotting, Western; Cell Line; DNA - genetics; DNA - isolation & purification; DNA, Antisense - genetics; DNA, Antisense - pharmacology; Dogs; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Isoenzymes - genetics; Isoenzymes - metabolism; Kidney; Kinetics; Protein Kinase C - genetics; Protein Kinase C - metabolism; RNA - genetics; RNA - isolation & purification; Tetradecanoylphorbol Acetate - pharmacology; Transfection; Transferases; Signal transduction; Protein kinase C; Enzyme; Inhibition; Antisense DNA; Arachidonic acid; Gene expression; Release
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)50291-5

A major unresolved issue in the area of signal transduction relates to the role of particular isoforms of protein kinase C (PKC) in mediating cellular responses subsequent to activation of that enzyme. We have addressed this issue by the use of antisense technology. We have stably transfected Madin-Darby canine kidney cells with antisense PKC alpha, PKC beta, or both PKC alpha and -beta cDNAs. The transfected cDNA was integrated and expressed. We have isolated cells in which expression of PKC alpha is inhibited. In cells transfected with antisense PKC alpha or both PKC alpha and -beta, phorbol ester-stimulated release of arachidonate and its metabolites was inhibited, whereas in cells transfected with antisense PKC beta cDNA alone, phorbol ester-stimulated arachidonate release was not significantly different from control cells. We thus demonstrate the use of a novel technique to inhibit PKC isoform expression. We show that inhibition of expression of PKC alpha causes a loss in phospholipase A2-mediated arachidonate release. Antisense-inhibited expression of PKC isoforms may provide a useful approach to define additional functions of particular PKC isoforms.