Is Polyamine Decrease a Common Feature of Apoptosis? Evidence from γ Rays- and Heat Shock-Induced Cell Death

Grassilli, Emanuela; Desiderio, Maria Alfonsina; Bellesia, Enrica; Salomoni, Paolo; Benatti, Francesca; Franceschi, Claudio

doi:10.1006/bbrc.1995.2679

Cited by 47 publications

(28 citation statements)

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“…1, shaded bars), as has been reported in some cells (36,37). A decline in putrescine content was seen in control and putrescine-treated cells, but the significance of this finding is hard to assess as putrescine is a relatively minor polyamine (note scale differences in Fig.…”

Section: Resultssupporting

confidence: 51%

Polyamine Regulation of Plasma Membrane Phospholipid Flip-Flop during Apoptosis

Bratton

Fadok

Richter

et al. 1999

Journal of Biological Chemistry

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During apoptosis, phosphatidylserine (PS) is moved from the plasma membrane inner leaflet to the outer leaflet where it triggers recognition and phagocytosis of the apoptotic cell. Although the mechanisms of PS appearance during apoptosis are not well understood, it is thought that declining activity of the aminophospholipid translocase and calcium-mediated, nonspecific flip-flop of phospholipids play a role. As previous studies in the erythrocyte ghost have shown that polyamines can alter flip-flop of phospholipids, we asked whether alterations in cellular polyamines in intact cells undergoing apoptosis would affect PS appearance, either by altering aminophospholipid translocase activity or phospholipid flip-flop. Cells of the human leukemic cell line, HL-60, were incubated with or without the ornithine decarboxylase inhibitor, difluoromethylornithine (DFMO), and induced to undergo apoptosis by ultraviolet irradiation. Whereas DFMO treatment resulted in profound depletion of putrescine and spermidine (but not spermine), it had no effect on caspase activity, DNA fragmentation, or plasma membrane vesiculation, typical characteristics of apoptosis. Notably, DFMO treatment prior to ultraviolet irradiation did not alter the decline in PS inward movement by the aminophospholipid translocase as measured by the uptake of 6-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)aminocaproyl] (NBD)-labeled PS detected in the flow cytometer. Conversely, the appearance of endogenous PS in the plasma membrane outer leaflet detected with fluorescein isothiocyanatelabeled annexin V and enhanced phospholipid flip-flop detected by the uptake of 1-palmitoyl-1-[6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)aminocaproyl]-sn-glycero-3-phosphocholine (NBD-PC) seen during apoptosis were significantly inhibited by prior DFMO treatment. Importantly, replenishment of spermidine, by treatment with exogenous putrescine to bypass the metabolic blockade by DFMO, restored both enhanced phospholipid flip-flop and appearance of PS during apoptosis. Such restoration was seen even in the presence of cycloheximide but was not seen when polyamines were added externally just prior to assay. Taken together, these data show that intracellular polyamines can modulate PS appearance resulting from nonspecific flip-flop of phospholipids across the plasma membrane during apoptosis.Under normal conditions, plasma membrane phospholipids of cells appear to be asymmetrically distributed across the bilayer with phosphatidylserine found almost entirely in the inner leaflet and sphingomyelin in the outer leaflet of the bilayer (1-3) . Maintenance of this asymmetry is attributed largely to the activity of the aminophospholipid translocase that moves phosphatidylserine (PS) 1 and, with a lesser affinity, phosphatidylethanolamine from the outer to the inner leaflet (4 -7). On the other hand, the appearance of PS, seen as a universal feature of apoptosis (3,8,9), and phospholipid flipflop during inflammatory cell activation (10 -13) require enhanced, nonspecific (with regard to head group) tra...

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Section: Resultssupporting

confidence: 51%

Polyamine Regulation of Plasma Membrane Phospholipid Flip-Flop during Apoptosis

Bratton

Fadok

Richter

et al. 1999

Journal of Biological Chemistry

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“…Initial studies on apoptosis in thymocytes observed polyamine depletion as an unsuspected finding and hypothesized that polyamine depletion contributed to the activation of apoptosis (25). Agmatine depletes intracellular polyamine levels to suppress growth (4,16,22,29,46,59,68).…”

Section: Discussionmentioning

confidence: 99%

“…As apoptotic models have implicated that polyamine depletion is a common and potentially causal factor of apoptosis (12,15,25,47), we hypothesized that the induction of apoptosis by agmatine may contribute to the antigrowth effects through cellular attrition. Apoptosis occurs during normal development and aging as a compensatory mechanism to cellular proliferation in maintaining cell populations.…”

mentioning

confidence: 99%

The arginine metabolite agmatine protects mitochondrial function and confers resistance to cellular apoptosis

Arndt¹,

Battaglia²,

Parisi³

et al. 2009

American Journal of Physiology-Cell Physiology

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“…This was thrown in doubt when it was established that the cells did not proliferate when ornithine was added in place of arginine. Replacement of arginine by citrulline, on the other hand, worked well and also prevented the development of hyperammonia (Van Rijn et al, 2003).The accumulation of ornithine in the medium and a report on the induction of ornithine decarboxylase by ionising radiation (Grassilli et al, 1995) pointed to the possibility that polyamines were involved in the formation of ammonia. Furthermore, since the medium toxicity was not fully explained by the presence of ammonia and cell death appeared to be apoptotic (Van Rijn et al, 2003), the idea was advanced that the cytotoxicity was partly due to disturbance of polyamine homeostasis (Schipper et al, 2000).…”

mentioning

confidence: 98%

“…The accumulation of ornithine in the medium and a report on the induction of ornithine decarboxylase by ionising radiation (Grassilli et al, 1995) pointed to the possibility that polyamines were involved in the formation of ammonia. Furthermore, since the medium toxicity was not fully explained by the presence of ammonia and cell death appeared to be apoptotic (Van Rijn et al, 2003), the idea was advanced that the cytotoxicity was partly due to disturbance of polyamine homeostasis (Schipper et al, 2000).…”

mentioning

confidence: 99%

Induction of hyperammonia in irradiated hepatoma cells: a recapitulation and possible explanation of the phenomenon

Rijn

Berg

Schipper³

et al. 2004

Br J Cancer

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We recently reported that a high amount of ammonia was produced in the culture medium of H35 hepatoma cells in reponse to ionising radiation (Van Rijn et al, 2003). As a result of hyperammonia and the accompanying pH increase, surviving cells failed to develop into colonies at high inoculation densities. The expected clonogenic potential of the cells reverted to more normal levels when the culture medium of irradiated cultures was replaced by a fresh medium halfway through the postexposure period. Even so, colonies were reduced in size. The removed medium proved cytotoxic to nonirradiated cells, indicating that irradiation per se was not responsible. Analysis of the medium implicated ammonia as the primary cytotoxic compound. Endogenously produced ammonia gas acts like a free radical, and is very aggressive and cytotoxic until it is converted into ammonium ions, which cells can tolerate at millimolar levels. Since native ammonia produces alkalinity, raised pH also accounts for cell death. Parallel with the development of hyperammonia, ornithine and citrulline levels were raised, whereas arginine was lost from the medium. Thus, this minimal deviation hepatoma seemed to be able to convert ammonia into urea through a fully functional ornithine cycle. This was thrown in doubt when it was established that the cells did not proliferate when ornithine was added in place of arginine. Replacement of arginine by citrulline, on the other hand, worked well and also prevented the development of hyperammonia (Van Rijn et al, 2003).The accumulation of ornithine in the medium and a report on the induction of ornithine decarboxylase by ionising radiation (Grassilli et al, 1995) pointed to the possibility that polyamines were involved in the formation of ammonia. Furthermore, since the medium toxicity was not fully explained by the presence of ammonia and cell death appeared to be apoptotic (Van Rijn et al, 2003), the idea was advanced that the cytotoxicity was partly due to disturbance of polyamine homeostasis (Schipper et al, 2000). An excess of polyamines then could be a potential source for the ammonia (Seiler, 2000).Investigation of the synthesis and interconversion pathways of the diamine, putrescine, and the polyamines, spermidine and spermine was undertaken. Several inhibitors are known to interfere with the key enzymes, ornithine decarboxylase and Sadenosylmethionine decarboxylase, or the diamine-and polyamine oxidases. Many of the inhibitors, for example, a-difluoromethylornithine, L-agmatine, dicyclohexylamine, methylglyoxalbis-guanylhydrazone and CGP48664A, reduced the release of ammonia. Conversely, the addition of the depleted product (putrescine, spermidine or spermine) or of inhibitors of the diamine/polyamine oxidases (aminoguanidine and MDL72527) restored hyperammonia. However, any suppression of the hyperammonia development was accompanied by a decreased conversion of arginine into ornithine and citrulline. Furthermore, most of the inhibitors except L-agmatine reduced the clonogenic survival in the combination ...

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Is Polyamine Decrease a Common Feature of Apoptosis? Evidence from γ Rays- and Heat Shock-Induced Cell Death

Cited by 47 publications

References 0 publications

Polyamine Regulation of Plasma Membrane Phospholipid Flip-Flop during Apoptosis

Polyamine Regulation of Plasma Membrane Phospholipid Flip-Flop during Apoptosis

The arginine metabolite agmatine protects mitochondrial function and confers resistance to cellular apoptosis

Induction of hyperammonia in irradiated hepatoma cells: a recapitulation and possible explanation of the phenomenon

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