Effect of polyamine depletion on caspase activation: a study with spermine synthase-deficient cells.

Activation of the caspase proteases represents a central point in apoptosis. The requirement for spermine for the processes leading to caspase activation has been studied in transformed embryonic fibroblasts obtained from gyro (Gy) mutant male mice. These cells lack spermine synthase activity and thus provide a valuable model to study the role of spermine in cell processes. Gy fibroblasts do not contain spermine and have a higher spermidine content. However, when compared with fibroblasts obtained from normal male littermates (N cells), Gy fibroblasts were observed to grow normally. The lack of spermine did not affect the expression of Bcl-2, and caspases 3 and 9 were activated by etoposide in both N and Gy cells, indicating that spermine is dispensable for caspase activation. Spermine deficiency did not significantly influence caspase activity in cells treated with etoposide, cycloheximide or staurosporine, but sensitized the cells to UV irradiation, which triggered significantly higher caspase activity in Gy cells compared with N cells. alpha-Difluoromethylornithine (DFMO), an inhibitor of polyamine synthesis that is able to deplete cells of putrescine and spermidine, but usually does not influence spermine content, was able to produce a more complete polyamine depletion in Gy cells. This depletion, which included spermine deficiency, dramatically increased caspase activation and cell death in Gy fibroblasts exposed to UV irradiation. On the other hand, in either N or Gy cells, DFMO treatment did not influence caspase activity triggered by staurosporine, but inhibited it when the inducers were cycloheximide or etoposide. In Gy cells depleted of polyamines by DFMO, polyamine replenishment with either spermidine or spermine was sufficient to restore caspase activity induced by etoposide, indicating that, in this model, polyamines have an interchangeable role in supporting caspase activation. Therefore, spermine is not required for such activation, and the effect and specificity of polyamine depletion on caspase activity may be very different, depending on the role of polyamines in the specific death pathways engaged by different stimuli. Some inducers of apoptosis, for example etoposide, absolutely require polyamines for caspase activation, yet the lack of polyamines, particularly spermine, strongly increases caspase activation when induced by UV irradiation.

[1]  K. Abromeit.  Music Received , 2022, Notes.

[2]  J. Nilsson,et al.  Skin fibroblasts from spermine synthase-deficient hemizygous gyro male (Gy/Y) mice overproduce spermidine and exhibit increased resistance to oxidative stress but decreased resistance to UV irradiation. , 2000, The Biochemical journal.

[3]  A. Pegg,et al.  Effect of spermine synthase deficiency on polyamine biosynthesis and content in mice and embryonic fibroblasts, and the sensitivity of fibroblasts to 1,3-bis-(2-chloroethyl)-N-nitrosourea. , 2000, The Biochemical journal.

[4]  Tak W. Mak,et al.  Two Distinct Pathways Leading to Nuclear Apoptosis , 2000, The Journal of experimental medicine.

[5]  C. Caldarera,et al.  Polyamines directly induce release of cytochrome c from heart mitochondria. , 2000, Biochemical Journal.

[6]  W. Earnshaw,et al.  Induction of apoptosis by cancer chemotherapy. , 2000, Experimental cell research.

[7]  J. Barrett,et al.  Regulation of p53 stability and activity in response to genotoxic stress. , 2000, Mutation research.

[8]  A. Strasser,et al.  Apoptosis and cell division. , 2000, Current opinion in cell biology.

[9]  J. Meyer,et al.  Effect of Age on the Expression of Pex (Phex) in the Mouse , 2000, Calcified Tissue International.

[10]  P. Harari,et al.  Modulation of tumor cell proliferation and apoptosis by polyamine depletion in cells of head and neck squamous cell carcinomas. , 1999, Radiation research.

[11]  H. Steller,et al.  Death by design: mechanism and control of apoptosis. , 1999, Trends in cell biology.

[12]  V. Fadok,et al.  Polyamine Regulation of Plasma Membrane Phospholipid Flip-Flop during Apoptosis* , 1999, The Journal of Biological Chemistry.

[13]  S. Oredsson,et al.  Topoisomerase II is nonfunctional in polyamine‐depleted cells , 1999, Journal of cellular biochemistry.

[14]  C. Caldarera,et al.  Spermine triggers the activation of caspase‐3 in a cell‐free model of apoptosis , 1999, FEBS letters.

[15]  C. Franceschi,et al.  2-deoxy-d-ribose-induced apoptosis in HL-60 cells is associated with the cell cycle progression by spermidine. , 1999, Biochemical and biophysical research communications.

[16]  L. Fong,et al.  α-Difluoromethylornithine Inhibits N-Nitrosomethylbenzylamine-induced Esophageal Carcinogenesis in Zinc-deficient Rats: Effects on Esophageal Cell Proliferation and Apoptosis , 1998 .

[17]  C. Caldarera,et al.  Spermine causes caspase activation in leukaemia cells , 1998, FEBS letters.

[18]  S. Kaufmann.  Cell death induced by topoisomerase-targeted drugs: more questions than answers. , 1998, Biochimica et biophysica acta.

[19]  N. Osheroff,et al.  Mechanism of action of eukaryotic topoisomerase II and drugs targeted to the enzyme. , 1998, Biochimica et biophysica acta.

[20]  G. Evan,et al.  A matter of life and cell death. , 1998, Science.

[21]  S. Cory,et al.  The Bcl-2 protein family: arbiters of cell survival. , 1998, Science.

[22]  P. Gregersen,et al.  Cerebral Ischemia Enhances Polyamine Oxidation: Identification of Enzymatically Formed 3-Aminopropanal as an Endogenous Mediator of Neuronal and Glial Cell Death , 1998, The Journal of experimental medicine.

[23]  C. Caldarera,et al.  Inhibition of etoposide-induced apoptosis with peptide aldehyde inhibitors of proteasome. , 1998, The Biochemical journal.

[24]  P. Vandenabeele,et al.  Sensitization of tnf-induced apoptosis with polyamine synthesis inhibitors in different human and murine tumour cell lines. , 1998, Cytokine.

[25]  T. Meitinger,et al.  Spermine deficiency in Gy mice caused by deletion of the spermine synthase gene. , 1998, Human molecular genetics.

[26]  C. Franceschi,et al.  Polyamine depletion protects HL-60 cells from 2-deoxy-D-ribose-induced apoptosis. , 1998, Life sciences.

[27]  N. Thornberry,et al.  A Combinatorial Approach Defines Specificities of Members of the Caspase Family and Granzyme B , 1997, The Journal of Biological Chemistry.

[28]  P. De Feudis,et al.  Treatment with inhibitors of polyamine biosynthesis, which selectively lower intracellular spermine, does not affect the activity of alkylating agents but antagonizes the cytotoxicity of DNA topoisomerase II inhibitors. , 1997, British Journal of Cancer.

[29]  E. Gerner,et al.  Loss of intracellular putrescine pool-size regulation induces apoptosis. , 1997, Experimental cell research.

[30]  A. Pegg,et al.  Induction of apoptosis by excessive polyamine accumulation in ornithine decarboxylase-overproducing L1210 cells. , 1995, The Biochemical journal.

[31]  C. Kahana,et al.  Exposure to ornithine results in excessive accumulation of putrescine and apoptotic cell death in ornithine decarboxylase overproducing mouse myeloma cells. , 1995, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[32]  J. Cleveland,et al.  Ornithine decarboxylase is a mediator of c-Myc-induced apoptosis , 1994, Molecular and cellular biology.

[33]  L. Dillehay,et al.  The effect of polyamine depletion on the cytotoxic response to PUVA, gamma rays and UVC in V79 cells in vitro. , 1994, Biochemical and biophysical research communications.

[34]  A. Pegg,et al.  Spermidine/spermine N1‐acetyltransferase — the turning point in polyamine metabolism , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[35]  A. Pegg,et al.  Enhancement of the spermidine uptake system and lethal effects of spermidine overaccumulation in ornithine decarboxylase-overproducing L1210 cells under hyposmotic stress. , 1993, The Journal of biological chemistry.

[36]  A. U. Khan,et al.  A proposed function for spermine and spermidine: protection of replicating DNA against damage by singlet oxygen. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[37]  J. L. Mitchell,et al.  Abnormal accumulation and toxicity of polyamines in a difluoromethylornithine-resistant HTC cell variant. , 1992, Biochimica et biophysica acta.

[38]  V. Brunton,et al.  Mechanisms of spermine toxicity in baby-hamster kidney (BHK) cells. The role of amine oxidases and oxidative stress. , 1991, The Biochemical journal.

[39]  B. Feuerstein,et al.  Implications and concepts of polyamine‐nucleic acid interactions , 1991, Journal of cellular biochemistry.

[40]  Pierce Gb,et al.  Polyamine oxidation, programmed cell death, and regulation of melanoma in the murine embryonic limb. , 1989 .

[41]  A. Pegg,et al.  Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. , 1988, Cancer research.

[42]  L. Zwelling,et al.  Effect of polyamine depletion by alpha-difluoromethylornithine or (2R,5R)-6-heptyne-2,5-diamine on drug-induced topoisomerase II-mediated DNA cleavage and cytotoxicity in human and murine leukemia cells. , 1987, Cancer research.

[43]  M. Lyon,et al.  The Gy mutation: another cause of X-linked hypophosphatemia in mouse. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[44]  W. Gahl,et al.  Reversal by aminoguanidine of the inhibition of proliferation of human fibroblasts by spermidine and spermine. , 1978, Chemico-biological interactions.

[45]  M. Drezner.  PHEX gene and hypophosphatemia. , 2000, Kidney international.

[46]  T. Efferth,et al.  Impact of viral oncogenesis on responses to anti-cancer drugs and irradiation. , 2000, Critical reviews in oncogenesis.

[47]  H. Wallace,et al.  Changes in polyamine catabolism in HL-60 human promyelogenous leukaemic cells in response to etoposide-induced apoptosis. , 1999, The Biochemical journal.

[48]  S H Kaufmann,et al.  Mammalian caspases: structure, activation, substrates, and functions during apoptosis. , 1999, Annual review of biochemistry.

[49]  Y. Yano,et al.  The antiproliferative effect of HGF on hepatoma cells involves induction of apoptosis with increase in intracellular polyamine concentration levels. , 1998, Oncology reports.

[50]  L. Fong,et al.  Alpha-difluoromethylornithine inhibits N-nitrosomethylbenzylamine-induced esophageal carcinogenesis in zinc-deficient rats: effects on esophageal cell proliferation and apoptosis. , 1998, Cancer research.

[51]  E. Løvaas.  Antioxidative and metal-chelating effects of polyamines. , 1997, Advances in pharmacology.

[52]  R. Madhubala,et al.  Difluoromethylornithine antagonizes taxol cytotoxicity in MCF-7 human breast cancer cells. , 1997, Oncology research.

[53]  G. B. Pierce,et al.  Polyamine oxidation, programmed cell death, and regulation of melanoma in the murine embryonic limb. , 1989, Cancer research.

[54]  L. Marton,et al.  Effects of treatment with DNA-directed cancer chemotherapeutic agents after polyamine depletion. , 1987, Pharmacology & therapeutics.

[55]  C. Stefanelli,et al.  Separation of N1- and N8-acetylspermidine isomers by reversed-phase column liquid chromatography after derivatization with dansyl chloride. , 1986, Journal of chromatography.