Fermented Wheat Germ Extract Inhibits Glycolysis/Pentose Cycle Enzymes and Induces Apoptosis through Poly(ADP-ribose) Polymerase Activation in Jurkat T-cell Leukemia Tumor Cells

• Published in: The Journal of biological chemistry 2002-11, Vol.277 (48), p.46408-46414
• Main Authors: Comin-Anduix, Begona, Boros, Laszlo G, Marin, Silvia, Boren, Joan, Callol-Massot, Carles, Centelles, Josep J, Torres, Josep L, Agell, Neus, Bassilian, Sara, Cascante, Marta
• Format: Article
• Language: English
• Subjects: Apoptosis - drug effects; Carbon Isotopes; Enzyme Activation; Enzyme Inhibitors - pharmacology; Fermentation; Gas Chromatography-Mass Spectrometry; Glucosephosphate Dehydrogenase - antagonists & inhibitors; Glucosephosphate Dehydrogenase - metabolism; Glycolysis; Hexokinase - antagonists & inhibitors; Hexokinase - metabolism; Humans; Jurkat Cells; L-Lactate Dehydrogenase - antagonists & inhibitors; L-Lactate Dehydrogenase - metabolism; Lactates - metabolism; Leukemia, T-Cell - enzymology; Leukemia, T-Cell - pathology; Pentose Phosphate Pathway; Plant Extracts - pharmacology; Poly(ADP-ribose) Polymerases - metabolism; Transketolase - antagonists & inhibitors; Transketolase - metabolism; Triticum - embryology; Triticum aestivum
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M206150200

The fermented extract of wheat germ, trade name Avemar, is a complex mixture of biologically active molecules with potent anti-metastatic activities in various human malignancies. Here we report the effect of Avemar on Jurkat leukemia cell viability, proliferation, cell cycle distribution, apoptosis, and the activity of key glycolytic/pentose cycle enzymes that control carbon flow for nucleic acid synthesis. The cytotoxic IC 50 concentration of Avemar for Jurkat tumor cells is 0.2 mg/ml, and increasing doses of the crude powder inhibit Jurkat cell proliferation in a dose-dependent fashion. At concentrations higher than 0.2 mg/ml, Avemar inhibits cell growth by more than 50% (72 h of incubation), which is preceded by the appearance of a sub-G 1 peak on flow histograms at 48 h. Laser scanning cytometry of propidium iodide- and annexin V-stained cells indicated that the growth-inhibiting effect of Avemar was consistent with a strong induction of apoptosis. Inhibition by benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone of apoptosis but increased proteolysis of poly(ADP-ribose) indicate caspases mediate the cellular effects of Avemar. Activities of glucose-6-phosphate dehydrogenase and transketolase were inhibited in a dose-dependent fashion, which correlated with decreased 13 C incorporation and pentose cycle substrate flow into RNA ribose. This decrease in pentose cycle enzyme activities and carbon flow toward nucleic acid precursor synthesis provide the mechanistic understanding of the cell growth-controlling and apoptosis-inducing effects of fermented wheat germ. Avemar exhibits about a 50-fold higher IC 50 (10.02 mg/ml) for peripheral blood lymphocytes to induce a biological response, which provides the broad therapeutic window for this supplemental cancer treatment modality with no toxic effects.