Induction of apoptotic cell death by putrescine.
The polyamines are essential for cellular growth and differentiation. Ornithine decarboxylase (ODC), which catalyses the first step in the biosynthesis of the polyamines, has a very fast turnover and is subject to a strong feedback control by the polyamines. In the present study, we show that overexpression of a metabolically stable ODC in CHO cells induced a massive cell death unless the cells were grown in the presence of the ODC inhibitor alpha-difluoromethylornithine (DFMO). Cells overexpressing wild-type (unstable) ODC, on the other hand, were not dependent on the presence of DFMO for their growth. The induction of cell death was correlated with a dramatic increase in cellular putrescine levels. Analysis using flow cytometry revealed perturbed cell cycle kinetics, with a large accumulation of cells with sub-G1 amounts of DNA, which is a typical sign of apoptosis. Another strong indication of apoptosis was the finding that one of the key enzymes in the apoptotic process, caspase-3, was induced when DFMO was omitted from the growth medium. Furthermore, inhibition of the caspase activity significantly reduced the recruitment of cells to the sub-G1 fraction. In conclusion, deregulation of polyamine homeostasis may negatively affect cell proliferation and eventually lead to cell death by apoptosis if putrescine levels become too high.
Takao
Koichi
K
Department of Experimental Medical Science, Lund University, S-22184 Lund, Sweden.
Rickhag
Mattias
M
Hegardt
Cecilia
C
Oredsson
Stina
S
Persson
Lo
L
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D016466:CHO Cells; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D000518:Eflornithine / Q000494:pharmacology; D004791:Enzyme Inhibitors; D016193:G1 Phase / Q000187:drug effects; D009955:Ornithine Decarboxylase / Q000378:metabolism; D065108:Ornithine Decarboxylase Inhibitors; D011700:Putrescine / Q000096:biosynthesis / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D065108:Ornithine Decarboxylase Inhibitors; D053148:Caspase 3; D020169:Caspases; D009955:Ornithine Decarboxylase; D011700:Putrescine; D000518:Eflornithine