Protein-kinase-C iso-enzymes support DNA synthesis and cell survival in colorectal-tumor cells.
Protein-kinase-C signalling has been blocked in colorectal tumor cells by kinase inhibitors, by TPA down-regulation or by exposure to anti-sense oligonucleotides. This resulted in growth inhibition in all cell lines used. The kinase inhibitors H7 and calphostin induced apoptosis, demonstrated by the appearance of cells with characteristically condensed chromatin and the induction of stand-breaks in the DNA. A cell-death-inducing concentration of 15 microgram/ml H7 down-regulated the bcl-2 levels after 9 hr, while bak levels were not affected. Gö6976,-an inhibitor of Ca(++)-dependent PKC iso-enzymes, was not active in growth inhibition or induction of apoptosis. Analysis of DNA synthesis in inhibitor-treated cultures indicated that H7 caused strong inhibition in all cell lines, while the more specific inhibitor calphostin was effective only in VACO235 adenoma cells. When down-regulation by TPA or anti-sense oligonucleotides was used to block PKC, effects on cell numbers were smaller and delayed. However, induction of apoptosis was significantly increased in SW480 carcinoma cells 4 days after exposure to anti-epsilon and anti-zeta oligonucleotides in SW480 and T84 carcinoma cells. Apoptosis was preceeded by loss of PKC protein and of bcl-2 from day 1 after addition of the oligonucleotides. In VACO235 adenoma cells, no induction of apoptosis could be observed when anti-epsilon and anti-zeta oligonucleotides were used. On the other hand, the adenoma cells were more responsive to anti-alpha and anti-beta oligonucleotides, which strongly inhibited DNA-synthesis 3 days after addition to the culture medium. Our results indicate that the Ca(++)-dependent PKCs alpha and beta are involved in proliferation signals, while the Ca(++)-independent PKCs epsilon and zeta are involved in survival pathways of colorectal tumor cells.
Hochegger
K
K
Institute of Tumor Biology/Cancer Research, University of Vienna, Vienna, Austria.
Partik
G
G
Schörkhuber
M
M
Marian
B
B
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D017209:Apoptosis; D002273:Carcinogens / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000502:physiology*; D004273:DNA, Neoplasm / Q000096:biosynthesis*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019073:HT29 Cells / Q000187:drug effects / Q000473:pathology; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000502:physiology*; D008565:Membrane Proteins / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology; D050998:bcl-2 Homologous Antagonist-Killer Protein
C494772:BAK1 protein, human; D002273:Carcinogens; D004273:DNA, Neoplasm; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D008565:Membrane Proteins; D009281:Naphthalenes; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D050998:bcl-2 Homologous Antagonist-Killer Protein; C061465:calphostin complex; C080501:1-(5-isoquinolinylsulfonyl)-3-methylpiperazine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate