Metabolic and antiproliferative consequences of activated polyamine catabolism in LNCaP prostate carcinoma cells.
Depletion of intracellular polyamine pools invariably inhibits cell growth. Although this is usually accomplished by inhibiting polyamine biosynthesis, we reasoned that this might be more effectively achieved by activation of polyamine catabolism at the level of spermidine/spermine N(1)-acetyltransferase (SSAT); a strategy first validated in MCF-7 breast carcinoma cells. We now examine the possibility that, due to unique aspects of polyamine homeostasis in the prostate gland, tumor cells derived from it may be particularly sensitive to activated polyamine catabolism. Thus, SSAT was conditionally overexpressed in LNCaP prostate carcinoma cells via a tetracycline-regulatable (Tet-off) system. Tetracycline removal resulted in a rapid approximately 10-fold increase in SSAT mRNA and an increase of approximately 20-fold in enzyme activity. SSAT products N(1)-acetylspermidine, N(1)-acetylspermine, and N(1),N(12)-diacetylspermine accumulated intracellularly and extracellularly. SSAT induction also led to a growth inhibition that was not accompanied by polyamine pool depletion as it was in MCF-7 cells. Rather, intracellular spermidine and spermine pools were maintained at or above control levels by a robust compensatory increase in ornithine decarboxylase and S-adenosylmethionine decarboxylase activities. This, in turn, gave rise to a high rate of metabolic flux through both the biosynthetic and catabolic arms of polyamine metabolism. Treatment with the biosynthesis inhibitor alpha-difluoromethylornithine during tetracycline removal interrupted flux and prevented growth inhibition. Thus, flux-induced growth inhibition appears to derive from overaccumulation of metabolic products and/or from depletion of metabolic precursors. Metabolic effects that were not excluded as possible contributing factors include high levels of putrescine and acetylated polyamines, a 50% reduction in S-adenosylmethionine, and a 45% decline in the SSAT cofactor acetyl-CoA. Overall, the study demonstrates that activation of polyamine catabolism in LNCaP cells elicits a compensatory increase in polyamine biosynthesis and downstream metabolic events that culminate in growth inhibition.
Kee
Kristin
K
Grace Cancer Drug Center, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Vujcic
Slavoljub
S
Merali
Salim
S
Diegelman
Paula
P
Kisiel
Nicholas
N
Powell
C Thomas
CT
Kramer
Debora L
DL
Porter
Carl W
CW
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D012437:Adenosylmethionine Decarboxylase / Q000378:metabolism; D002277:Carcinoma / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D003839:Deoxyadenosines / Q000378:metabolism; D000518:Eflornithine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D008297:Male; D008715:Methionine / Q000494:pharmacology; D009952:Ornithine / Q000494:pharmacology; D009955:Ornithine Decarboxylase / Q000378:metabolism; D065108:Ornithine Decarboxylase Inhibitors; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors; D011073:Polyamines / Q000737:chemistry / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D011700:Putrescine / Q000031:analogs & derivatives* / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives / Q000378:metabolism; D013754:Tetracyclines / Q000494:pharmacology; D013872:Thionucleosides / Q000378:metabolism; D014407:Tumor Cells, Cultured; D000097667:Polyamine Oxidase; D000099228:Diamine N-Acetyltransferase
D000105:Acetyl Coenzyme A; D000123:Acetyltransferases; D012437:Adenosylmethionine Decarboxylase; D003839:Deoxyadenosines; D000518:Eflornithine; D004791:Enzyme Inhibitors; D008715:Methionine; D009952:Ornithine; D009955:Ornithine Decarboxylase; D065108:Ornithine Decarboxylase Inhibitors; D000587:Oxidoreductases Acting on CH-NH Group Donors; D011073:Polyamines; D011700:Putrescine; D012333:RNA, Messenger; D012436:S-Adenosylmethionine; D013754:Tetracyclines; D013872:Thionucleosides; D000097667:Polyamine Oxidase; D000099228:Diamine N-Acetyltransferase; C044445:MDL 72527; C008500:5'-methylthioadenosine