Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “polyamines”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 541 records · Page 30Linked to original sources

Lack of a correlation between polyamine synthesis and DNA synthesis by cultured rat liver cells and fibroblasts.

Growth stimulation of either fetal rat liver cells or rat embryo fibroblasts in culture results in considerable increases in intracellular polyamine levels as cells proceed through the cell cycle. Treatment of such cell cultures with appropriate levels of two inhibitors of polyamine synthesis, namely alpha-hydrazino ornithine and methylglyoxal bis (guanylhydrazone), can essentially completely block these increases in cellular polyamine content. Under such conditions, where the elevation in intracellular polyamine content is prevented, cell cultures are nevertheless able to initiate DNA synthesis and subsequently synthesize DNA at rates comparable to untreated control cultures that have been growth-stimulated. These two cell types therefore contain sufficient polyamines when in a resting state (G1) to enable them to enter from G1 into S phase and traverse S phase at normal rates in the absense of further polyamine synthesis. The recruitment of cells into the first cell cycle, through serum stimulation of growth, therefore appears not to be mediated or regulated by the increases in intracellular levels of polyamines that occurs under these conditions. Conversely, the arrest of growth of these cell types resulting from serum deprivation is not mediated by a limitation of intracellular polyamine content.

Blood↗

Effects of polyamines on cyclic AMP-mediated stimulation of amino acid transport in isolated rat hepatocytes.

The effects of natural polyamines on cyclic AMP-mediated stimulation of amino acid transport in isolated rat hepatocytes were analyzed. Despite the fact that polyamines could directly compete with alpha-aminoisobutyric acid (AIB) for uptake, preincubation of hepatocytes with polyamines did not significantly alter basal AIB transport. The stimulatory effect of glucagon or cyclic AMP analogs was differently affected by polyamines, since it was reduced in the presence of spermine and, inversely, potentiated by spermidine, putrescine, and cadaverine. Dose-dependence analysis showed that half maximal and maximal effects occurred with 2-3 and 6-10 mM external concentrations, respectively. None of the polyamine effects could be ascribed to transstimulation or transinhibition of amino acid uptake. The inhibitory effect exerted by spermine correlated its capacity to inhibit [3H]-leucine incorporation into proteins partially. The potentiating effect of the other polyamines did not result from stabilization of newly synthesized carrier proteins. Instead, the increase in Vmax of the high affinity transport component suggested that more carriers became available, presumably because polyamines facilitated their synthesis by interacting directly with one or several steps controlled by cyclic AMP. Polyamines appear to represent a new class of factors capable of modulating the cyclic AMP-mediated stimulation of amino acid transport, in hepatocytes.

Aminoisobutyric Acids↗

Role of polyamines in the stimulation of synthesis and secretion of plasminogen activator from bovine aortic endothelial cells.

The effects of the polyamines putrescine (PUT), spermidine (SPD), and spermidine (SPM) on the secretion of plasminogen activator (PA) and plasminogen activator inhibitor (PAI) were evaluated using cultured bovine aortic endothelial cells. All three polyamines enhanced PA secretion in a time- and dose-dependent manner, with a potency rank order of SPM greater than SPD greater than PUT. The PA stimulation required both RNA and protein synthesis, as evidenced by inhibition of polyamine-induced PA secretion by actinomycin D and cycloheximide. The inhibitors of polyamine biosynthesis methylglyoxal bis-(guanylhydrazone) (MGBG) and dl-(difluoromethyl) ornithine (DFMO) alone did not affect basal or polyamine-induced PA secretion, with the exception that MGBG reduced the effect of PUT. Polyamine-treated cells enhanced secretions of both tissue-type and urokinase-type PA. The results of the present study suggest that polyamines may play a role in the regulation of PA synthesis and secretion and that this function can be modified under pathophysiological conditions affecting cellular and tissue levels of polyamines.

Animals↗

EGF stimulates polyamine uptake in Caco-2 cells.

We investigated the effects of epidermal growth factor (EGF) on polyamine uptake in Caco-2 cell monolayers. Cells were grown until confluence (day 7) or until differentiation (day 14). Polyamine uptake into Caco-2 cells was stimulated by EGF in a dose-dependent manner. Both basal and EGF-stimulated uptake rates were higher in 7- than in 14-day-old Caco-2 cells. Stimulation with EGF resulted in a significant increase in Vmax and an increased affinity for putrescine and spermine. Polyamine uptake was not inhibited when protein synthesis was blocked by cycloheximide, implying that no additional protein synthesis occured for stimulatory effect of EGF on polyamine uptake. The tyrosine kinase inhibitor, genistein, completely inhibited EGF-stimulated polyamine uptake, indicating that tyrosine phosphorylation plays a role in EGF-stimulated polyamine uptake in Caco-2 cells. The effect of EGF on polyamine uptake into Caco-2 cells, therefore, could be due to translocation of intracellular proteins which were not previously incorporated into the membrane, or direct alteration of polyamine transporter.

Adenocarcinoma↗

The expression of Escherichia coli SOS genes recA and uvrA is inducible by polyamines.

Polyamines are polycationic compounds that have been implicated in a variety of cellular processes. We first report that the expression of recA and uvrA genes in Escherichia coli is inducible by polyamines. The recA and uvrA genes were effectively inducible by spermine (tetra-amine) and less effectively inducible by spermidine (tri-amine). On the other hand, both genes were not significantly inducible by putrescine (diamine) and divalent cation Mg(2+). The expression of both genes was dependent on the charge of the polyamine in the order of spermine (+4), spermidine (+3), and putrescine (+2). The induction of recA and uvrA genes by polyamines showed a dose-dependent relationship and no synergistic effects. Introduction of gyrA mutation conferring DNA relaxation increased the basal expression of recA gene about 2.5-fold compared to the wild type, but did not significantly affect the polyamine-dependent induction ratios of the recA gene. The basal and the polyamine-dependent expression of uvrA gene are not dependent on gyrA mutation. These results suggest that the polyamine-dependent expression of recA and uvrA genes may be regulated in a different way. Our results indicated that polyamines are involved in the SOS induction of recA and uvrA genes at transcriptional levels in E. coli.

Adenosine Triphosphatases↗

Novel lysine-spermine conjugate inhibits polyamine transport and inhibits cell growth when given with DFMO.

Polyamines are ubiquitous molecules with multiple intracellular functions. Cells tightly regulate their levels through feedback mechanisms affecting synthesis, intracellular conversion, and transport. Because polyamines have an important role in regulating cell growth, they are a target for cancer therapeutic development. However, to effectively inhibit cell growth through polyamine depletion one needs to inhibit both polyamine synthesis and import. Although the mammalian polyamine transporter has not been cloned, we have identified ORI 1202, an N(1)-spermine-L-lysinyl amide, as an effective polyamine transport inhibitor. ORI 1202 prevents the cellular accumulation of [(3)H]spermidine over a 20-h test period. ORI 1202 (30-100 microM) effectively inhibits cell growth when used in conjunction with the polyamine synthesis inhibitor alpha-difluoromethylornithine (DFMO; > or =230 microM). Human breast, prostate, and bladder carcinoma cell lines and melanoma cell lines show ORI 1202 EC(50) values in the low micromolar range when tested in conjunction with DFMO. This cytostatic effect correlates with a reduction in the intracellular levels of putrescine and spermidine. When ORI 1202 (45 mg/kg, i.p., tidx5) and DFMO (1% in drinking water) were delivered over 14 days, MDA-MB-231 breast tumor xenografts in nude mice showed 50% growth inhibition. Polyamine depletion therapy provides a cytostatic therapy that could be useful against cancer and other diseases resulting from uncontrolled cell growth.

Animals↗

Involvement of polyamines in apoptosis. Facts and controversies: effectors or protectors?

The natural polyamines (putrescine, spermidine and spermine) are ubiquitous low-molecular aliphatic amines that play multifunctional roles in cell growth and differentiation. Recently, evidence has merging that polyamines are actively involved in cell death. Changes in polyamine homeostasis have been reported during cell death of nerve cells, in programmed cell death of embryonic cells and in various in vitro models of apoptosis. Polyamines and many of their structural analogs exert cytotoxic effects in vitro as well in vivo. Furthermore, polyamine analogs and inhibitors of the polyamine anabolic/catabolic pathways modulate processes of cell death in a cell-type specific way. Much ambiguity exists in the working mechanisms by which polyamines mediate apoptosis since they have been shown to act as promoting, modulating or protective agents in apoptosis. Nevertheless, from the studies reviewed here it can be concluded that polyamines are critically involved in cellular survival which makes them suitable targets for therapeutic intervention that is specifically directed to cell death pathways.

Animals↗

In vivo, synergestic inhibition of MAT-LyLu rat prostatic adenocarcinoma growth by polyamine deprivation and low-dose cyclophosphamide.

Polyamine deprivation in vivo produces significant tumor growth inhibition of the hormone-resistant, metastatic Dunning Mat-LyLu murine prostatic carcinoma. In order to produce a cytotoxic effect in addition to the cytostatic effect of polyamine deprivation, various chemotherapy regimens, combined with drug-containing polyamine-deficient chow (DC-PDC), were assessed. Triple chemotherapy combining methotrexate, cyclophosphamide and vindesine; and monochemotherapy with high-dose cyclophosphamide (90 mg. kg-1) and low-dose cyclophosphamide (20 mg.kg-1) were studied alone and in combination with DC-PDC. A variant of DC-PDC excluding the polyamine oxidase inhibitor MDL 72527 was also studied in combination with low-dose cyclophosphamide. The triple-chemotherapy regimen alone or in combination with polyamine deprivation was effective on tumor growth inhibition but was also toxic. High-dose cyclophosphamide alone produced significant tumor growth inhibition and an increase in life span. High-dose cyclophosphamide in combination with DC-PDC was also effective on tumor growth but was also toxic. Low-dose cyclophosphamide alone was moderately effective on tumor growth inhibition with a marginal increase in life span. When combined with polyamine deprivation, results with low-dose cyclophosphamide compared favourably with those of high-dose cyclophosphamide alone and prevented the formation of lung metastases. The polyamine oxidase inhibitor does not appear to be mandatory to achieve this effect if DC-PDC is combined with low-dose cyclophosphamide. Polyamine deprivation appears to be an important tool in anticancer therapy, allowing the use of reduced doses of cytotoxic agents with the same antitumoral efficacy.

Adenocarcinoma↗

Polyamine levels of human colorectal adenocarcinomas are correlated with tumor stage and grade.

BACKGROUND AND AIMS: Cellular proliferation and differentiation are regulated by polyamines and their rate-limiting enzyme ornithine decarboxylase (ODC), both of which are correlated with tumor growth, but their role in differentiation is less clear. We investigated the correlation of ODC activity and polyamine levels with tumor stage and grade with respect to sample recruitment. PATIENTS AND METHODS: We determined ODC activity ([(14)C]CO(2) release), polyamines (HPLC), and histological staging and grading (TNM classification) of tissue samples from 64 patients with colorectal adenocarcinomas. RESULTS: We found the concentrations of putrescine, spermidine, and N(1)-acetyl-spermidine and the ODC activity in tumor tissue to be twice as high as in adjacent normal mucosa. A critical parameter affecting ODC activity was ischemic time, which significantly reduced ODC activity levels in tumors (threefold) and in the surrounding normal tissue (ninefold) when the ischemic period exceeded 1 h. By contrast, polyamine content was not affected by ischemia. Total polyamine and spermine concentrations were higher in T3 and T4 than in T2 tumors, but putrescine was higher in T4 than in T3 and T2 tumors. There were significantly higher levels of total polyamines and spermine in moderately differentiated (G2) than in poorly differentiated (G3) tumors. CONCLUSION: The lower spermidine/spermine ratio in G2 (0.44) compared with that in G3 (0.64) tumors suggests the involvement of the polyamines in colonic cell differentiation. Polyamine content is thus correlated with the tumor stage.

Acetyltransferases↗

Ectomycorrhizal fungal species and strains differ in their ability to produce free and conjugated polyamines.

Production of free and conjugated polyamines by one strain of Laccaria proxima (Boud.) Maire, three strains (H, O, K) of Paxillus involutus (Batsch) Fr., and one strain of Pisolithus tinctorius was studied in vitro. Spermidine (Spd) was the main polyamine in the 4-week-old mycelium of all the fungi. It was mainly present in the free form, but it also occurred in conjugated forms. Paxillus involutus strain H released large amounts of free putrescine (Put), and the Pisolithus tinctorius released a compound probably related to cadaverine (Cad). On the other hand, these two fungi contained less conjugated polyamines than the other fungi. In addition to the amounts, the forms (perchloric acid soluble and insoluble) of conjugated polyamines in the mycelium varied between species and strains. L. proxima contained nearly as much insoluble conjugated Spd as free Spd, whereas Paxillus involutus strains O and K contained relatively large amounts of soluble conjugated Spd. The results suggest that ectomycorrhizal fungal species and strains differ in their ability and need to produce conjugated polyamines. The small amounts of soluble conjugated polyamines found in the culture filtrates indicate that some specific conjugated polyamines may be involved in polyamine translocation across the plasma membrane.

Agaricales↗

Polyamines are absorbed through a y+ amino acid carrier in rat intestinal epithelial cells.

Due to the similarity in transport characteristics of polyamines and the y+ basic amino acid system, we hypothesized that both substrates could be moving through a common carrier site. Competitive and cross inhibition experiments in intestinal epithelial cells revealed the possibility of a common transport site. N-ethylmalemide (NEM) inhibited both lysine and putrescine transport, confirming that both were carried by a y+ transporter. Overexpressing the y+ transporter CAT-1 in a polyamine transport-deficient cell line, CHO-MG, did not reconstitute polyamine-transport. Thus, polyamines are not traveling through CAT-1. To determine if lysine is carried by a polyamine transport site, an antizyme-overexpressing cell line was used. Antizyme overexpression decreased polyamine uptake by 50%; in contrast, lysine transport was unaffected. Therefore, lysine is not traveling through a polyamine transport site. It appears that polyamines and lysine are likely traveling through a common unknown y+ transport site.

Absorption↗

Polyamine transport in Neurospora crassa.

Polyamine transport in Neurospora crassa is concentrative and energy dependent in a dilute buffer. The saturable systems governing the uptake of putrescine (Km = 0.6 mM), spermidine (Km = ca. 0.24 mM), and spermine (Km = 0.07 mM) share components, as indicated by mutual inhibition among the polyamines. In addition, nonsaturable components prevail for putrescine and spermidine, particularly the former. Radiolabeled substrates, once in the cell, are released only slowly, even if unlabeled polyamines are included in the incubation medium. Permeabilization of cells with n-butanol leads to partial release of internalized 14C-polyamines, and the remainder is almost wholly exchangeable with added, unlabeled polyamine. Polyamine uptake was inhibited by the polyamines themselves and by a polyamine analog, methylglyoxal bisguanylhydrazone, but only weakly and incompletely by the basic amino acids arginine and ornithine. Uptake of putrescine and spermidine was inhibited by monovalent cations, Ca2+, and certain other components of the growth medium. As a result, uptake from the growth medium was very slow and largely by way of the nonsaturable uptake mechanism.

Biological Transport↗

Fluorescence polarization studies on Escherichia coli membrane stability and its relation to the resistance of the cell to freeze-thawing. II. Stabilization of the membranes by polyamines.

The effects of polyamines, spermine, spermidine and putrescine on the stabilization of the membrane organization of Escherichia coli cells were studied using measurements of fluorescence polarization change of extrinsic fluorescence probes in membrane specimens as a function of temperature. The effects of the polyamines on the restoration of the cell viability after freeze-thawing were also investigated. In logarithmic-phase membrane specimens, polyamines depressed the polarization ratio increase below the transition temperatures in a dose-dependent manner. The physiologically relevant concentration of polyamines repressed the ratios to the same levels as are obtained with the stationary-phase specimens. In the stationary-phase specimens, no effect of polyamines on repression of the polarization increase was observed. A preliminary exposure of logarithmic-phase cells to polyamines protected the cells from the reduction of viability in freeze-thawing. However, a considerably high concentration and a certain length of preincubation time were required in order to an effect to be exerted. These results indicate that the intracellular polyamines could stabilize the membrane organization of logarithmic-phase cells to the same extent as in the stationary-phase cell membranes. It is conjectured that the membrane stability which is mediated by the polyamines results in cellular resistance to freeze-thawing, as it is attained by increasing the growth phase of the cells.

Cell Membrane↗

Inhibition of polyamine accumulation and cell proliferation by derivatives of diaminopropane in Ehrlich ascites cells grown in culture.

1. 1,3-Diaminopropane and some of its derivatives are potent inhibitors of ornithine decarboxylase (EC 4.1.1.17) in Ehrlich ascites cells grown in suspension culture. Among the amine derivatives tested, 1,3-diamino-2-propanol most effectively prevented any accumulation of spermidine and spermine in ascites cells when the proliferation was stimulated by diluting the cells with fresh medium. 2. The effectiveness of diaminopropanol in abolishing polyamine accumulation was primarily based on a rapid decay of ornithine decarboxylase activity following the exposure of the cells to the drug. 3. The mechanism of action of diaminopropanol on ornithine decarboxylase apparently involved a formation of macromolecular inhibitors or 'antizymes' to the enzyme. 4. Even though the inhibitory effect of 1,3-diaminopropane on polyamine accumulation approached that of diaminopropanol, the former compound only marginally inhibited the incorporation of [3H]thymidine into DNA and that of [14C]leucine into protein, in contrast to the marked depression of macromolecular synthesis produced by diaminopropanol. The apparent dissociation of polyamine depletion brought about by 1,3-diaminopropane from an antiproliferative action was apparently due to the fact that diaminopropane, unlike diaminopropanol, was partially capable of taking over the function of natural polyamines. 5. The inhibition of DNA and protein synthesis as well as the prevention of increase in cell number by diaminopropanol was closely associated with polyamine depletion and was fully comparable, as regards timing and magnitude, with that achieved with difluoromethylornithine. The antiproliferative effect of diaminopropanol, however, was only partly reversed by a simultaneous addition of putrescine (or spermidine) into the culture medium. The lack of a complete reversal of the action of diaminopropanol on cell growth by natural polyamines was apparently due to the fact that it was remarkably difficult or even impossible to increase intracellular polyamine concentrations by exogenous polyamines in the presence of diaminopropanol. Nevertheless, the diaminopropanol-induced arrest of growth was reversible as judged by a rapid increase in ornithine decarboxylase activity followed by restoration of DNA synthesis.

Adenosylmethionine Decarboxylase↗

Dietary restriction, polyamines and monocrotaline-induced pulmonary hypertension.

Dietary restriction (DR), i.e. reduction of total caloric intake, has been shown to result in protection against monocrotaline (MCT)-induced pulmonary hypertension (PH). Restriction of the diet to 8 g/rat/day instead of the usual intake (18 g/rat/day), inhibits the progression of cardiopulmonary changes and prolongs survival after a single dose of MCT. We have shown previously that the development of MCT-induced pulmonary hypertension is associated with inhibition of polyamine biosynthesis in the lungs of MCT-treated rats. In the present study, we tested the hypothesis that DR provides protection against the development of chronic PH in the rat by limiting increases in polyamine and DNA synthesis. We randomly divided animals into four groups each (MCT, MCT + DR, control, and control + DR). We injected rats with a single dose of MCT (60 mg/kg, s.c.) and a corresponding number of control rats with vehicle. Animals in all groups were given free access to food and water prior to administration of MCT. Immediately following injection of MCT both the MCT and control groups were given free access to food and water, while the other groups (MCT + DR and control + DR) we given the restricted diet (8 g/rat/day). Daily measurements were made of body weight and of water and food intake. Animals were killed in each group at 1, 4, 7, 14, and 21 days post MCT to determine right ventricular hypertrophy (RVH), lung wet weight, ornithine decarboxylase (ODC) activity, and polyamine and DNA contents. We measured DNA synthesis 7 days after MCT by determining [3H]thymidine incorporation into the whole lung DNA. We found that 7 days after MCT treatment DNA synthesis increased compared to control. However, DR (MCT + DR) treatmen prevented the increase in DNA synthesis following MCT. Right ventricular hypertrophy, lung wet weight, ODC activity and lung polyamine levels were increased following MCT. Treatment with DR (MCT + DR) prevented increases in RVH, lung wet weight, ODC activity and lung polyamine levels. We conclude that DR to 8 g/day/rat protects against MCT-induced PH and is associated with an inhibition of increased lung polyamine and DNA synthesis that occur in the lung during the development of MCT-induced PH. These results are consistent with a recent report which suggests that increased lung polyamine biosynthesis is required for the development of MCT-induced PH. The data are also consistent with the hypothesis that inhibition of polyamine biosynthesis influences the development of MCT-induced PH in part by regulating DNA synthesis in key lung cells.

Animals↗

Immunocytochemical localization of polyamines in the tiger salamander retina.

The polyamines spermine and spermidine are present in neural tissue, but their functions there are not well understood. Recent work suggests that the NMDA subtype of glutamate receptors, other glutamate receptor subtypes, and certain K(+)-channels, are neural targets for polyamines. To better understand the neuron-specific roles of polyamines, we have developed antibodies that interact with spermine and spermidine in aldehyde-fixed tissue and used these antibodies in immunocytochemical studies to determine the cellular localization of these polyamines in the tiger salamander retina. The affinity-purified, polyclonal antibodies were highly specific for spermine and spermidine, exhibiting < 1% cross reactivity with putrescine, and virtually no cross-reactivity with GABA, arginine, lysine, or glutaraldehyde. Polyamine labeling was most abundant in cells in the inner half of the inner nuclear layer and in the ganglion cell layer. Some cells in the outer half of the inner nuclear layer are labeled, and there was some labeling in both synaptic layers. Double-labeling experiments indicated (1) all GABAergic amacrine cells were polyamine-positive; and (2) all ganglion cells (identified by back-filling after microinjections of rhodamine in the optic nerve) were polyamine-positive. These results are consistent with a role for polyamines as modulators of NMDA receptor function and channel function in the inner retina.

Animals↗

Protein-bound polyamines in the plasma of mice grafted with the Lewis lung carcinoma.

Protein-bound polyamines were isolated from the plasma of mice using antipolyamine antibodies covalently linked to magnetic latex spheres. Their subsequent separation by polyacrylamide gel electrophoresis (PAGE) showed that in plasma from normal mice, 3 proteins (27, 55 and 82 kDa) carrying polyamines could be visualized, whereas in mice bearing the Lewis lung carcinoma at least 8 other proteins of higher molecular mass (5 of 94, 110, 130, 145 and 160 kDa, and 3 of greater than 170 kDa) had bound polyamines. These protein-bound polyamines could be detected from the first week after tumour graft; they increased during the second and third week but decreased thereafter. These proteins were not bound by immunolatex spheres preincubated with spermine bound to a protein-carrier insulin. Moreover, the appearance of these protein-bound polyamines was not a consequence of the inflammatory process since in mice infected with heat-inactivated Brucella abortus, with the exception of a 65 kDa protein, polyamines were bound to the same proteins found in normal mice. In mice grafted with the Lewis lung carcinoma the concomitant decrease in transglutaminase-mediated polyamine (e.g. putrescine) binding capacity of plasma proteins provides additional evidence for the presence in vivo of polyamines already bound to plasma proteins.

Animals↗

Polyamine depletion of cells reduces the infectivity of herpes simplex virus but not the infectivity of Sindbis virus.

The effect of polyamines on the viral growth was examined using cell strains that could be effectively depleted of polyamines. In order to avoid the polyamines present in serum we used a polyamine auxotrophic Chinese hamster ovary cell line P22 growing in serum-free medium and Vero cells growing in low serum medium. The final yield of an enveloped RNA virus, Sindbis, in P22 cells was not decreased by depletion of cellular polyamines although the onset of the viral replication was delayed. In contrast the final yield of an enveloped DNA virus, Herpes simplex virus (HSV), was considerably reduced in Vero cells, depleted of polyamines by alpha-difluoromethylornithine, an inhibitor of polyamine synthesis. However, the number of HSV particles detected by electronmicroscopy was not decreased. Southern blot analysis of HSV-DNA from the polyamine depleted and the control cells showed changes in the relative abundance of the DNA fragments suggesting that impairment in DNA synthesis may have caused the decreased infectivity of HSV.

Animals↗