ABSTRACT
Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis and a target for chemoprevention. Hydroxydibenzoylmethane (HDB), a derivative of dibenzoylmethane of licorice, is a promising chemopreventive agent. In this paper, we investigated whether HDB would inhibit the ODC pathway to enhance apoptosis in human promyelocytic leukemia HL-60 cells. We found ODC enzyme activity was reduced during HDB treatment. Overexpression of ODC in HL-60 parental cells could reduce HDB-induced apoptosis, which leads to loss of mitochondrial membrane potential (Deltapsim), through lessening intracellular ROS. Furthermore, ODC overexpression protected cytochrome c release and the activation of caspase-3 following HDB treatment. The results demonstrated HDB-induced apoptosis was through a mechanism of down-regulation of ODC and occurred along a ROS-dependent mitochondria-mediated pathway.
Subject(s)
Humans , Apoptosis/drug effects , Caspase 3/metabolism , Chalcones/metabolism , Chemoprevention , Cytochromes c/biosynthesis , Down-Regulation , Gene Expression , HL-60 Cells , Immunoblotting , Leukemia, Myeloid/enzymology , Membrane Potential, Mitochondrial/drug effects , Mitochondria/enzymology , Ornithine Decarboxylase/antagonists & inhibitors , Reactive Oxygen Species/analysis , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Ovarian sizes (length and width) were measured in young females of Anastrepha fraterculus (Wiedemann) (Diptera, Tephritidae) subjected or not to the inhibitor α -difluormethylornithine (α -DFMO). The most effective concentration of α -DMFO used was 50 mM and the ovarian measurements (length and width) of the treated females were smaller than those of females not treated with α -DMFO. These data may suggest some relationship between ornithine decarboxylase (ODC) and sexual maturation in A. fraterculus.
As dimensões dos ovários (comprimento e largura) foram mensuradas em fêmeas jovens da Anastrepha fraterculus (Wiedemann) (Diptera, Tephritidae) submetidas ou não ao inibidor α -difluormetilornitina (α -DFMO). A concentração mais efetiva de α -DMFO utilizada foi 50 mM e as medidas (comprimento e largura) das fêmeas tratadas com o inibidor foram menores que as fêmeas não tratadas com inibidor α -DMFO. Estes dados podem sugerir uma relação entre ornitina descarboxilase (ODC) e maturação sexual em A. fraterculus.
Subject(s)
Animals , Female , Eflornithine/pharmacology , Enzyme Inhibitors/pharmacology , Ornithine Decarboxylase/antagonists & inhibitors , Ovary/drug effects , Tephritidae/drug effects , Organ Size/drug effects , Ovary/growth & development , Tephritidae/anatomy & histology , Tephritidae/enzymologyABSTRACT
The multiplication of A. culbertsoni in the peptone medium was not inhibited by 10-20 mM concentration of alpha-difluoromethyl ornithine (DMFO) while a partial and transient inhibition of cell multiplication was observed by 10-20 mM DFMO in proteose peptone, yeast extract, glucose (PYG) medium. Ornithine decarboxylase (ODC) activity in the cells and cell free extracts was strongly inhibited by DFMO, excluding enzyme refractoriness and impermeability of cells for DFMO as the possible causes of DFMO resistance. The presence of polyamines in the peptone and PYG media as well as uptake of polyamines by the amoebae has been demonstrated. The growth and multiplication of A. culbertsoni in chemically defined medium was not affected by 1-5 mM DFMO while 10-20 mM DMFO yielded partial inhibition. A lowering of diaminopropane levels and enhancement of spermidine levels was observed in DFMO inhibited cells and level of ODC was drastically reduced in the inhibited cultures. Uptake of polyamines from the growth media may partly account for DFMO resistance of A. culbertsoni. Alternative mechanisms for DFMO resistance are indicated.
Subject(s)
Acanthamoeba/drug effects , Animals , Eflornithine/pharmacology , Ornithine Decarboxylase/antagonists & inhibitorsABSTRACT
Effect of snuff extract (SE) on cell proliferation as measured by 3H thymidine (TdR) uptake, induction of ornithine decarboxylase (ODC) and aryl hydrocarbon hydroxylase (AHH) was studied in primary embryonal mouse tongue cultures. Cultures treated with SE in combination with 7,12-dimethylbenz(a)anthracene (DMBA) showed inhibition of cell proliferation and decrease of ODC and AHH activities, compared to control, DMBA, and DMBA + 12-O-tetradecanoylphorbol 13-acetate treated cultures.