Biphasic effect of protein kinase C activators on spontaneous and glucocorticoid-induced apoptosis in primary mouse thymocytes.
Spontaneous and glucocorticoid (fluocinolone acetonide, FA)-induced apoptosis of primary mouse thymocytes was inhibited by protein kinase C (PKC) activators such as bryostatin-1 and phorbol ester 12-O-tetradecanoyl-phorbol-13 acetate (TPA) within the first 2-4 h of incubation but was enhanced upon prolonged treatment. Only the anti-apoptotic but not the pro-apoptotic effect of TPA was completely suppressed by the PKC inhibitor Goe 6983 and moderately inhibited by Goe 6976. Immunoblot analysis revealed distinct PKC alpha, beta, delta, eta, theta, mu and zeta signals, a very faint PKCepsilon and no PKCgamma signal. Upon prolonged TPA treatment all PKC isoenzymes became downregulated, albeit at different rates (PKCdelta>alpha>mu>beta,theta>>eta,zeta). No significant generation of caspase-derived catalytic PKC fragments, as found to be produced upon induction of apoptosis and to be pro-apoptotic in other systems, was observed in FA- or TPA-treated thymocytes. It is concluded that the early anti-apoptotic effect of TPA depends on the activation of n-type PKC isoenzymes, whereas stimulation of spontaneous and FA-induced apoptosis by TPA ensues, at least partially, from a downregulation (or inactivation) of anti-apoptotic PKC species, i.e. in primary thymocytes PKC activation is primarily involved in a negative regulation of apoptosis.
Rennecke
J
J
Research Program Tumor Cell Regulation, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120, Heidelberg, Germany.
Richter
K H
KH
Häussermann
S
S
Stempka
L
L
Strand
S
S
Stöhr
M
M
Marks
F
F
D000818:Animals; D017209:Apoptosis; D054713:Bryostatins; D002227:Carbazoles / Q000494:pharmacology; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D015536:Down-Regulation; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005446:Fluocinolone Acetonide / Q000037:antagonists & inhibitors / Q000494:pharmacology; D005938:Glucocorticoids / Q000494:pharmacology*; D015151:Immunoblotting; D007211:Indoles / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D007783:Lactones / Q000494:pharmacology; D018942:Macrolides; D051379:Mice; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000037:antagonists & inhibitors / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology / Q000187:drug effects*
D054713:Bryostatins; D002227:Carbazoles; D005938:Glucocorticoids; D007211:Indoles; D007527:Isoenzymes; D007783:Lactones; D018942:Macrolides; D005446:Fluocinolone Acetonide; C081021:Go 6976; C046785:bryostatin 1; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate