Effects of human immune interferon on cell viability, (2',5')oligoadenylate synthesis, and polyamine-dependent protein phosphorylation in human colon carcinoma cells in vitro.
The growth-inhibitory effect of human immune interferon (IFN-gamma) was investigated in human colon carcinoma cell line HT-29. Three-day treatment of HT-29 cells with IFN-gamma (10 to 200 units/ml) resulted in 30 to 90% growth inhibition and 40 to 99% reduction in colony formation. Measurement of DNA, RNA, and protein synthesis following IFN-gamma treatment showed a dose-dependent reduction in all 3 parameters. The associated changes in (2',5')oligoadenylate [(2',5')oligo(A)] pathway were measured under growth-inhibitory conditions. Upon 1-day exposure to 25 to 200 units/ml of IFN-gamma, (2',5')oligo(A) synthetase activity was induced 10- to 15-fold and remained elevated for 3 days, whereas (2',5')oligo(A) phosphodiesterase activity remained unchanged. There was no detectable increase in intracellular (2',5')oligo(A) levels after IFN-gamma treatment, and ribosomal RNA degradation was not observed. Accompanying 1-day treatment with IFN-gamma (100 units/ml) was an induction of a polyamine-dependent protein kinase, which was double-stranded RNA-independent and phosphorylated endogenous polypeptides with molecular weights of 68,000 and 72,000. A similar exposure of cells to IFN-gamma (25 to 100 units/ml) resulted in 30 to 70% inhibition of ornithine decarboxylase activity; however, no significant alteration in intracellular polyamine levels was observed. These data suggest that IFN-gamma-dependent toxicity is not related to (2',5')oligo(A) activation of a latent endoribonuclease but is accompanied by protein phosphorylation, which is, in part, stimulated by exogenous polyamines.
D015088:2',5'-Oligoadenylate Synthetase / Q000378:metabolism; D000227:Adenine Nucleotides / Q000096:biosynthesis*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003110:Colonic Neoplasms / Q000503:physiopathology*; D004261:DNA Replication / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000633:toxicity*; D007700:Kinetics; D009363:Neoplasm Proteins / Q000378:metabolism*; D009841:Oligonucleotides / Q000096:biosynthesis*; D009843:Oligoribonucleotides / Q000096:biosynthesis*; D010766:Phosphorylation; D011073:Polyamines / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D014158:Transcription, Genetic / Q000187:drug effects
D000227:Adenine Nucleotides; D009363:Neoplasm Proteins; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D011073:Polyamines; C023505:2',5'-oligoadenylate; D007371:Interferon-gamma; D015088:2',5'-Oligoadenylate Synthetase