Anti-mitochondrial effects of bisethyl polyamines in mammalian cells.
The effects of three bisethyl polyamine analogs on mitochondrial structure and function were examined in human HeLa and L1210 murine leukemia cells. N, N' Bis-[3(ethylamino)-propyl]1-7- heptane diamine (BEPH), and its octane (BEPO), and butane (BESPM) derivative, were shown by electron microscopy and/or Rhodamine 123 uptake studies to alter the structural integrity of mitochondria when both cell lines were treated at the approximate IC50 dose of each drug. At this dose, BEPH had no marked effects on levels of the naturally occurring polyamines, putrescine, spermidine or spermine, in either cell line whereas BEPO and BESPM treatment did result in pool depletion. Southern blot analysis demonstrated a time and dose-dependent loss of mitochondrial DNA from BEPH-treated L1210 cultures suggesting that loss of mitochondrial integrity extended to the DNA level. Treatment of L1210 cells with all three analogs revealed marked reductions in the activity of two mitochondrial enzymes citrate synthase and cytochrome C oxidase. HeLa cells treated with all three analogs exhibited markedly reduced levels of ATP, complete loss of cytidine triphosphate (CTP) and near total depletion of uridine triphosphate (CTP) and near total depletion of uridine triphosphate (UTP). There was also a loss of colony forming ability in HeLa cells which could be nearly completely reversed by the addition of either uridine or cytidine suggesting that NTP reduction may be the primary antiproliferative determinant in these cells. Growth inhibition by BEPH in L1210 cells was markedly potentiated by the glycolysis inhibitor, 2-deoxyglucose, which had no such effect in otherwise untreated cells. This suggests that BEPH treatment of L1210 cells results in impairment of mitochondrial ATP synthesis and activation of the glycolytic pathway for energy production. 2-deoxyglucose treatment also completely prevented the increase of ATP by BEPH treatment of L1210 cells. It is concluded that all three bisethyl polyamines alter HeLa and L1210 mitochondria both structurally and functionally and that these alterations may play a primary role in the antiproliferative activity of these agents in HeLa cells. In L1210, the different spectra of cellular biochemical changes following bisethyl polyamine treatment suggests that additional mechanisms may be in effect.
Marion Merrell Dow Research Institute, Cincinnati, Ohio 45215.
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Snyder
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Beach
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Loudy
D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000633:toxicity*; D015139:Blotting, Southern; D002455:Cell Division / Q000187:drug effects; D002950:Citrate (si)-Synthase / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000378:metabolism*; D003847:Deoxyglucose / Q000494:pharmacology; D003959:Diamines / Q000633:toxicity*; D003576:Electron Transport Complex IV / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D007939:Leukemia L1210; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D011073:Polyamines / Q000378:metabolism*; D012265:Ribonucleotides / Q000378:metabolism; D013096:Spermine / Q000031:analogs & derivatives* / Q000633:toxicity; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay; D000099228:Diamine N-Acetyltransferase
D000123:Acetyltransferases; D000970:Antineoplastic Agents; D002950:Citrate (si)-Synthase; D004272:DNA, Mitochondrial; D003847:Deoxyglucose; D003959:Diamines; D003576:Electron Transport Complex IV; D011073:Polyamines; D012265:Ribonucleotides; D013096:Spermine; D000099228:Diamine N-Acetyltransferase; C059685:N(1),N(11)-diethylnorspermine; C087842:N,N'-bis(3-(ethylamino)propyl)-1,8-octanediamine; C066203:N,N'-bis(3-(ethylamino)propyl)-1,7-heptanediamine