title: Folate deficiency-induced oxidative stress and apoptosis are mediated via homocysteine-dependent overproduction of hydrogen peroxide and enhanced activation of NF-kappaB in human Hep G2 cells.
abstract: Folate coenzymes are critical for de novo synthesis of purine and thymidine, and for interconversion of amino acids. Folate deficiency inhibits cellular proliferation, disturbs cell cycling, causes genetic damage and eventually results in cell death. Previously, we demonstrated that the demise of human hepatoma Hep G2 cells mediated by folate deficiency proceeded via a p53-independent apoptosis, and the perturbation of intracellular calcium homeostasis was also shown to be involved. To further delineate the mechanism associated with this observed phenomenon, Hep G2 cells were cultivated in the control or folate-deficient media (control media lacking folate, glycine, thymidine and hypoxanthine) for 4 weeks. At the end of this cultivation period, we found that TBARS (an index of lipid peroxidation) concentrations in the folate-deficient cells were drastically increased as compared to the control cells (0.04 vs 0.01 nmole/10(6) cells), indicating that a severe oxidative stress of the former cells had occurred. This phenomenon was also shown to coincide with the ability of these folate-deficient cells to elaborate increased amounts of H2O2 as compared to its folate-supplemented cells (2.87 vs 0.98 nmole/10(5) cells/h). Furthermore, the accelerated production of H2O2 by the folate-deficient cells was also closely correlated with the elevated homocysteine concentrations released in the culture medium (15.37 +/- 2.4 vs 3.58 +/- 2.4 micromole/L; P< 0.001). Finally, we demonstrated that folate deficiency was indeed capable of activating a redox-sensitive transcription factor, NF-kappaB, which is crucial in the control of a reactive oxygen species-mediated apoptosis. In summary, we show that folate deficiency-induced apoptosis is proceeded via the enhanced activation of NF-kappaB, which is the resulting form of the homocysteine-mediated overproduction of hydrogen peroxide.
authors.affiliation: Department of Biological Sciences, National Sun Yat-Shan University, Kaohsiung, Taiwan.
authors.forename: C L
authors.initials: CL
authors.lastname: Chern
authors.forename: R F
authors.initials: RF
authors.lastname: Huang
authors.forename: Y H
authors.initials: YH
authors.lastname: Chen
authors.forename: J T
authors.initials: JT
authors.lastname: Cheng
authors.forename: T Z
authors.initials: TZ
authors.lastname: Liu
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002374:Catalase / Q000096:biosynthesis; D002455:Cell Division / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004273:DNA, Neoplasm / Q000378:metabolism; D005492:Folic Acid / Q000494:pharmacology; D005494:Folic Acid Deficiency / Q000503:physiopathology*; D005979:Glutathione Peroxidase / Q000096:biosynthesis; D006710:Homocysteine / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008315:Malondialdehyde / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000502:physiology*; D014157:Transcription Factors; D014407:Tumor Cells, Cultured
chemical_list: D004273:DNA, Neoplasm; D016328:NF-kappa B; D014157:Transcription Factors; D006710:Homocysteine; D008315:Malondialdehyde; D005492:Folic Acid; D008715:Methionine; D006861:Hydrogen Peroxide; D002374:Catalase; D005979:Glutathione Peroxidase