Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-21T07:56:04.712Z Has data issue: false hasContentIssue false
  • English
  • Français

In vitro folate deficiency induces apoptosis by a p53, Fas (Apo-1, CD95) independent, bcl-2 related mechanism in phytohaemagglutinin-stimulated human peripheral blood lymphocytes

Published online by Cambridge University Press:  08 March 2007

Hui-Li Lin ,
Chung-Jern Chen ,
Wen-Chan Tsai ,
Jeng-Hsien Yen  and
Hong-Wen Liu
Hui-Li Lin
Affiliation:
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC
Chung-Jern Chen
Affiliation:
Division of Allergy, Immunology, RheumatologyDepartment of Internal MedicineCollege of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC
Wen-Chan Tsai
Affiliation:
Division of Allergy, Immunology, RheumatologyDepartment of Internal MedicineCollege of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC
Jeng-Hsien Yen
Affiliation:
Division of Allergy, Immunology, RheumatologyDepartment of Internal MedicineCollege of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC
Hong-Wen Liu*
Affiliation:
Division of Allergy, Immunology, RheumatologyDepartment of Internal MedicineCollege of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC
*
*Corresponding author: Dr Hong-Wen Liu, fax +886 7 3215842, email [email protected]
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

In vitro folate deficiency is associated with S phase accumulation and apoptosis in various cell types. To investigate the role of p53 and two apoptosis-related molecules, bcl-2 and Fas antigen (Apo-1, CD95), in the mechanism whereby folate-deficient lymphocytes accumulate and undergo apoptosis in the S phase, normal human peripheral blood lymphocytes were cultured for 3–9 d in control medium or in specially ordered and formulated HAM’ F-10 medium lacking folic acid, thymidine and hypoxanthine. Cells were stimulated with phytohaemagglutinin for the final 72 h prior to harvesting. The results indicate that p53 expression was downregulated in folate-deficient lymphocytes when compared with the control lymphocytes during the relevant period of S phase accumulation and apoptosis. In addition, folate deficiency was also found to downregulate IL-2, Fas antigen and bcl-2 expression, in terms of either mRNA or protein levels. The downregulation of Fas antigen suggests that folate deficiency-induced apoptosis probably does not occur via the Fas pathway. As IL-2 is a known inducer of bcl-2, and the downregulation of bcl-2 induces apoptosis, the downregulation of IL-2 and bcl-2 is suggested to play an important role in apoptosis. The complete rescue of folate-deficient lymphocytes from apoptosis was achieved by folic acid, thymidine or hypoxanthine alone or thymidine and hypoxanthine in combination. These results suggest that IL-2 depletion by folate deficiency in lymphocytes reduces the bcl-2 level, thereby triggering deoxynucleoside triphosphate pool imbalance and p53-independent apoptosis.

Keywords

Folate deficiencyApoptosisp53bcl-2
Type
Research Article
Information
British Journal of Nutrition , Volume 95 , Issue 5 , May 2006 , pp. 870 - 878
Copyright
Copyright © The Nutrition Society 2006

References

Benito, A, Silva, M, Grillot, D, Nunez, G & Fernandez-Luna, JLApoptosis induced by erythroid differentiation of human leukemia cell lines is inhibited by Bcl-XL. Blood 1996 87 3837–3843CrossRefGoogle ScholarPubMed
Benjamin, CB, Matthew, MM, Carol, MW, James, TMG, Robert, AH, Wang, GS, Wickramasinghe, N, Richard, BE & Ames, BNFolate deficiency causes uracil misincorporation into human DNA and chromosome breakage: implications for cancer and neuronal damage. Proc Natl Acad Sci USA 1997 94 3290–3295Google Scholar
Bunout, D, Petermann, M, Hirsch, S, de la Maza, P, Suazo, M, Barrera, G & Kauffman, RLow serum folate but normal homocysteine levels in patients with atherosclerotic vascular disease and matched healthy controls. Nutr 2000 16 434–438CrossRefGoogle ScholarPubMed
Collins, MK, Marvel, J, Malde, P & Lopez-Rivas, AInterleukin 3 protects murine bone marrow cells from apoptosis induced by DNA damaging agents. J Exp Med 1992 176 1043–1051CrossRefGoogle ScholarPubMed
Courtemanche, C, Huang, AC, Elson-Schwab, I, Kerry, N, Ng, BY & Ames, BNFolate deficiency and ionizing radiation cause DNA breaks in primary human lymphocytes: a comparison. FASEB J 2004 18 209–211CrossRefGoogle ScholarPubMed
Das, SK, Kunkel, TA & Loeb, LAEffects of altered nucleotide concentrations on the fidelity of DNA replication. Basic Life Sci 1985 31 117–126Google ScholarPubMed
Devireddy, LR & Green, MRTranscriptional program of apoptosis induction following interleukin 2 deprivation: identification of RC3, a calcium/calmodulin binding protein, as a novel proapoptotic factor. Mol Cell Biol 2003 23 4532–4541CrossRefGoogle ScholarPubMed
Donehower, LA & Bradley, AThe tumor suppressor p53. Biochim Biophys Acta 1993 1155 181–205Google ScholarPubMed
Duke, RC & Cohen, JJIL-2 addiction: withdrawal of growth factor activates a suicide program in dependent T cells. Lymphokine Res 1986 5 289–299Google ScholarPubMed
Duthie, SJ & Hawdon, ADNA instability (strand breakage, uracil misincorporation, and defective repair) is increased by folic acid depletion in human lymphocytes in vitro. FASEB J 1998 12 1491–1497CrossRefGoogle ScholarPubMed
Duthie, SJ, Narayanan, S, Brand, GM, Pirie, L & Grant, GImpact of folate deficiency on DNA stability. J Nutr 2002 132 S2444–S2449CrossRefGoogle ScholarPubMed
Fleischer, A, Ayllon, V, Dumoutier, L, Renauld, JC & Rebollo, AProapoptotic activity of ITM2B(s), a BH3-only protein induced upon IL-2-deprivation which interacts with Bcl-2. Oncogene 2002 21 3181–3189CrossRefGoogle ScholarPubMed
Fleming, A & Copp, AJEmbryonic folate metabolism and mouse neural tube defects. Science 1998 280 2107–2109CrossRefGoogle ScholarPubMed
Freudenheim, JL, Graham, S, Marshall, JR, Haughey, BP, Cholewinski, S & Wilkinson, GFolate intake and carcinogenesis of the colon and rectum. Int J Epidemiol 1991 20 368–374CrossRefGoogle ScholarPubMed
Hildeman, DA, Zhu, Y, Mitchell, TC, Kappler, J & Marrack, PMolecular mechanisms of activated T cell death in vivo. Curr Opin Immunol 2002 14 354–359CrossRefGoogle ScholarPubMed
Huang, RF, Ho, YH, Lin, HL, Wei, JS & Liu, TZFolate deficiency induces a cell cycle-specific apoptosis in HepG2 cells. J Nutr 1999 129 25–31CrossRefGoogle ScholarPubMed
Ingram, CF, Davidoff, AN, Marais, E, Sherman, GG & Mendelow, BVEvaluation of DNA analysis for evidence of apoptosis in megaloblastic anaemia. Br J Haematol 1997 96 576–583CrossRefGoogle ScholarPubMed
James, SJ, Miller, BJ, McGarrity, LJ & Morris, SMThe effect of folic acid deficiency on deoxyribonucleotide pools and cell cycle distribution in mitogen-stimulated lymphocytes. Cell Prolif 1994 27 395–406CrossRefGoogle Scholar
James, SJ, Pogribny, IP, Pogribna, M, Miller, BJ, Jernigan, S & Melnyk, SMechanisms of DNA damage, DNA hypomethylation, and tumor progression in the folate/methyl-deficient rat model of hepatocarcinogenesis. J Nutr 2003 133 S3740–S3747CrossRefGoogle ScholarPubMed
Kim, YI, Shirwadkar, S & Choi, SW, Puchyr, M, Wang, Y & Mason, JBEffects of dietary folate on DNA strand breaks within mutation-prone exons of the p53 gene in rat colon. Gastroenterology 2000 119 151–161CrossRefGoogle ScholarPubMed
Koury, MJ, Price, JO & Hicks, GGApoptosis in megaloblastic anemia occurs during DNA synthesis by a p53-independent, nucleoside-reversible mechanism. Blood 2000 96 3249–3255CrossRefGoogle ScholarPubMed
Kuerbitz, SJ, Plunkett, BS, Walsh, WV & Kastan, MBWild-type p53 is a cell cycle checkpoint determinant following irradiation. Proc Natl Acad Sci USA 1992 89 7491–7495CrossRefGoogle ScholarPubMed
Lagergren, J & Reichard, PPurine deoxyribonucleosides counteract effects of hydroxyurea on deoxyribonucleoside triphosphate pools and DNA synthesis. Biochem Pharmacol 1987 36 2985–2991CrossRefGoogle ScholarPubMed
Liao, WT, Chang, KL, Yu, CL, Chen, GS, Chang, LW & Yu, HSArsenic induces human keratinocyte apoptosis by the Fas/Fas ligand pathway, which correlates with alterations in nuclear factor-Kb and activator protein-1 activity. J Invest Dermatol 2004 122 125–129CrossRefGoogle Scholar
Linke, SP, Clarkin, KC, Di Leonardo, A, Tsou, A & Wahl, GMA reversible, p53-dependent G0/G1 cell cycle arrest induced by ribonucleotide depletion in the absence of detectable DNA damage. Genes Dev 1996 10 934–947CrossRefGoogle ScholarPubMed
Lowe, SW, Schmitt, EM, Smith, SW, Osborne, BA & Jacks, Tp53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 1993 362 847–849CrossRefGoogle ScholarPubMed
Maltzman, W & Czyzyk, LUV irradiation stimulates levels of p53 cellular tumor antigen in nontransformed mouse cells. Mol Cell Biol 1984 4 1689–1694Google ScholarPubMed
Mashiyama, ST, Courtemanche, C, Elson-Schwab, I, Crott, J, Lee, BL, Ong, CN, Fenech, M & Ames, BNUracil in DNA, determined by an improved assay, is increased when deoxynucleosides are added to folate-deficient cultured human lymphocytes. Anal Biochem 2004 330 58–69CrossRefGoogle ScholarPubMed
Michalis, LK, Pappas, K, Tweddel, A, Kolaitis, N, Katsouras, CS, Loukas, S, Goudevenos, J & Bourantas, KSideris, DARelatively low red cell folate levels and acute coronary syndromes. Coron Artery Dis 2001 12 665–668CrossRefGoogle ScholarPubMed
Miyashita, T & Reed, JCbcl-2 gene transfer increases relative resistance of S49.1 and WEHI7.2 lymphoid cells to cell death and DNA fragmentation induced by glucocorticoids and multiple chemotherapeutic drugs Cancer Res 1992 52 5407–5411Google ScholarPubMed
Miyawaki, T, Uehara, T, Nibu, R, Tsuji, T, Yachie, A, Yonehara, S & Taniguchi, NDifferential expression of apoptosis-related Fas antigen on lymphocyte subpopulations in human peripheral blood. J Immunol 1992 149 3753–3758CrossRefGoogle ScholarPubMed
Oliver, FJ, Collins, MK & Lopez-Rivas, ARegulation of the salvage pathway of deoxynucleotides synthesis in apoptosis induced by growth factor deprivation. Biochem J 1996 316 421–425CrossRefGoogle ScholarPubMed
Oliver, FJ, Collins, MK & Lopez-Rivas, AOverexpression of a heterologous thymidine kinase delays apoptosis induced by factor deprivation and inhibitors of deoxynucleotide metabolism. J Biol Chem 1997 272 10624–10630CrossRefGoogle ScholarPubMed
Oliver, FJ, Marvel, J, Collins, MK & Lopez-Rivas, ABcl-2 oncogene protects a bone marrow-derived pre-B-cell line from 5 fluor,2deoxyuridine-induced apoptosis Biochem Biophys Res Commun 1993 194 126–132CrossRefGoogle ScholarPubMed
Pogribny, IP, Basnakian, AG, Miller, BJ, Lopatina, NG, Poirier, LA & James, SJBreaks in genomic DNA and within the p53 gene are associated with hypomethylation in livers of folate/methyl-deficient rats. Cancer Res 1995 55 1894–1901Google ScholarPubMed
Rosenberg, IH & Mason, JBFolate, dysplasia, and cancer. Gastroenterology 1989 97 502–503CrossRefGoogle ScholarPubMed
Shane, B & Stokstad, ELVitamin B12-folate interrelationships. Annu Rev Nutr 1985 5 115–141CrossRefGoogle ScholarPubMed
Smithells, RW, Sheppard, S & Schorah, CJVitamin deficiencies and neural tube defects. Arch Dis Child 1976 51 944–950CrossRefGoogle ScholarPubMed
Strasser, A, Harris, AW, Jacks, T & Cory, SDNA damage can induce apoptosis in proliferating lymphoid cells via p53-independent mechanisms inhibitable by Bcl-2. Cell 1994 79 329–339CrossRefGoogle ScholarPubMed
van der Weyden, MB, Hayman, RJ, Rose, IS & Brumley, JFolatedeficient human lymphoblasts: changes in deoxynucleotide metabolism and thymidylate cycle activities. Eur J Haematol 1991 47 109–114CrossRefGoogle ScholarPubMed
Wickramasinghe, SN & Fida, SBone marrow cells from vitamin B12- and folate-deficient patients misincorporate uracil into DNA. Blood 1994 83 1656–1661CrossRefGoogle ScholarPubMed
Yoshioka, A, Tanaka, S, Hiraoka, O, Koyama, Y, Hirota, Y, Ayusawa, D, Seno, T, Garrett, C & Wataya, YDeoxyribonucleoside triphosphate imbalance. 5-Fluorodeoxyuridine-induced DNA double strand breaks in mouse FM3A cells and the mechanism of cell death. J Biol Chem 1987 262 8235–8241CrossRefGoogle ScholarPubMed
Yu, HS, Liao, WT, Chang, KL, Yu, CL & Chen, GSArsenic induces TNF-a release and tumor necrosis factor receptor-1 signaling in T-helper cell apoptosis. J Invest Dermatol 2002 119 812–819CrossRefGoogle Scholar
Zambetti, GP & Levine, AJA comparison of the biological activities of wild-type and mutant p53. FASEB J 1993 7 855–865CrossRefGoogle ScholarPubMed