Inhibitors of cGMP-dependent protein kinase block senescence induced by inactivation of T antigen in SV40-transformed immortal human fibroblasts.
Immortal human fibroblasts isolated following transfection with thermolabile simian virus 40 T antigen lost division potential upon shift up in temperature due to heat inactivation of the antigen. Such cells showed a concomitant change in the distribution of a mortality marker, mortalin, from a juxtanuclear cap like distribution of immortal cells to a uniform cytosolic distribution of mortal cells. We made an attempt to modulate the above inducible system of cellular senescence using various protein kinase inhibitors. Among the indolocarbazole type inhibitors tested, only KT5823, defined as a specific inhibitor of cGMP-dependent protein kinase, blocked the loss of division potential as determined by cell growth and colony forming ability. This inhibitor also prevented the above change in mortalin distribution due to temperature shift. In addition, the isoquinoline sulfonamide derivatives H8, H9, H88 and H89, all shown to inhibit cGMP-dependent protein kinase, suppressed the senescence. Inhibitors specific to other types of protein kinases, protein phosphatases or tyrosine kinases tested had no effect. Since there was no difference between the effective and non-effective inhibitors in their effects on cell cycle progression, cell cycle arrest by itself cannot account for the above phenomenon. These results suggest that a signaling pathway possibly mediated by cGMP-dependent protein kinase is involved in the induction of cellular senescence.
Fujii
M
M
Department of Cellular and Molecular Biology, Hiroshima University School of Medicine, Japan.
Ide
T
T
Wadhwa
R
R
Tahara
H
H
Kaul
S C
SC
Mitsui
Y
Y
Ogata
T
T
Oishi
M
M
Ayusawa
D
D
D000470:Alkaloids / Q000494:pharmacology*; D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis* / Q000502:physiology; D015415:Biomarkers / Q000032:analysis; D002227:Carbazoles; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure; D002471:Cell Transformation, Neoplastic; D016922:Cellular Senescence / Q000187:drug effects*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors*; D003513:Cycloheximide / Q000494:pharmacology; D005347:Fibroblasts; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles; D007546:Isoquinolines / Q000494:pharmacology*; D024101:Mitochondrial Proteins; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D015398:Signal Transduction; D013539:Simian virus 40 / Q000235:genetics*; D019311:Staurosporine; D013449:Sulfonamides / Q000494:pharmacology*
D000470:Alkaloids; D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D002227:Carbazoles; D018840:HSP70 Heat-Shock Proteins; C492001:HSPA9 protein, human; D006360:Heat-Shock Proteins; D007211:Indoles; D007546:Isoquinolines; D024101:Mitochondrial Proteins; D013449:Sulfonamides; C080352:mortalin; C073601:KT 5823; D003513:Cycloheximide; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D019311:Staurosporine