Abstract
Primary central nervous system lymphomas (PCNSL) frequently reveal genomic instability. We analysed different functional genetic variants affecting the folate and homocysteine metabolism important for DNA integrity in 31 PCNSL patients and 142 controls. We found significantly less carriers of the methionine synthase c.2756A>G (D919G) missense polymorphism among the patients (0.16 vs 0.42; odds ratio 0.26, CI95%: 0.09–0.74; P=0.005), suggesting a protective function of the G allele. These data stimulate further epidemiological and functional studies focusing on the role of homocysteine and folate metabolism in lymphoma tumorigenesis.
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Main
Primary central nervous system lymphomas (PCNSL) are highly malignant non-Hodgkin's lymphomas (NHL). The vast majority of them are of the diffuse large B-cell type (Montesinos-Rongen et al, 1999; DLBCL; Garter and Warnke, 2001). Genetic instability with characteristic chromosomal imbalances is a characteristic feature of PCNSL (Weber et al, 2000; Gonzalez-Gomez et al, 2003; Nakamura et al, 2003). Since homocysteine and folate metabolism is closely linked to DNA methylation, it considerably contributes to the maintenance of DNA integrity (Figure 1; Mudd et al, 2001). Consequently, genetic variants that functionally influence homocysteine and folate metabolism are associated with different types of cancer, for example, systemic NHL, acute leukaemia and colorectal cancer (Ma et al, 1997; Skibola et al, 1999; Matsuo et al, 2001; Gonzalez-Fraile et al, 2002; Lincz et al, 2003). To test the hypothesis that genetic variants of the homocysteine and folate metabolism alter the susceptibility of developing PCNSL, we analysed five polymorphisms affecting both the availability of 5,10-methylenetetrahydrofolate as a cosubstrate in nucleic acid synthesis and the level of S-adenosylmethionine (SAM) as a cosubstrate in DNA methylation.
Materials and methods
We performed a case–control study using DNA samples of 31 German patients with PCNSL of the DLBCL-subtype (m/f: 11/20; age: 61±10 years; Pels et al, 2003). DNA samples of 142 healthy German individuals (m/f: 62/80; age: 63±14 years), matched for the area of residence, served as controls. Genomic DNA was prepared according to standard methods (Miller et al, 1988). Five functional polymorphisms were analysed as previously described: Cystathionin beta-synthase (CBS) c.833T>C (I278 T) and 844ins68 (splice variant), methionine synthase (MS) c.2756A>G (D919G), and methylenetetrahydrofolate reductase (MTHFR) c.677C>T (A222 V) and c.1298A>C (E429A). With the exception of CBS I278 T, these polymorphisms have a high prevalence within the healthy German population with allele frequencies ranging between 0.08 and 0.34 (Harmon et al, 1999; Linnebank et al, 2000, 2001).
Logistic regression analysis was used to examine the association of single or combined polymorphisms with PCNSL and to exclude confounding effects of age, gender or multiple testing. In addition, χ2-tests were used to test differences of allele frequencies in patients and controls. Threshold was defined with P<0.05. The study was approved by the local ethics committee, and all participants gave written informed consent.
Results
The Hardy–Weinberg equilibrium was confirmed for all polymorphisms. Allele frequencies of the control group did not significantly differ from those reported in the literature (Harmon et al, 1999; Linnebank et al, 2000, 2001; Table 1). The MS c.2756A>G polymorphism (G-allele) was significantly less frequent in patients than in controls. In all, 6% of controls, but none of the patients were homozygous for the G-allele. At least one mutant allele was detected in 16% of PCNSL patients compared to 42% of controls (odds ratio (OR): 0.26, CI95%: 0.09 – 0.74; P=0.005 in regression analysis; P=0.007 in uncorrected χ2-analysis; P=0.035 in χ2-analysis corrected for multiple testing).
The CBS 844ins68 polymorphism was detected more frequently in patients (26%) than in controls (14%). However, the difference did not reach statistical significance in regression analysis (OR: 2.43, CI95%: 0.90 – 6.61; P=0.08) or χ2-analysis (P=0.09, uncorrected). Further, we did not find any significant association of the other polymorphisms (MTHFR c.677C>T or c.1298A>C, CBS c.833T>C) or combinations with PCNSL.
Discussion
In the present study, we investigated the association of functional variants of the homocysteine and folate metabolism with PCNSL. Previous studies found that the MS missense dimorphism c.2756A>G (D919G) is associated with a lower risk of developing colorectal cancer and types of systemic NHL (Ma et al, 1999; Lincz et al, 2003). Our findings suggest a protective function of MS c.2756A>G against PCNSL.
Methionine synthase c.2756A>G reduces MS activity (Harmon et al, 1999) leading to a lower consumption of 5,10- methylenetetrahydrofolate by the homocysteine metabolism. As a consequence, increased levels of 5,10-methylenetetrahydrofolate are available for nucleic acid synthesis, thus reducing the risk of thymidin deprivation and uracil misincorporation associated with chromosomal breakage (Blount et al, 1997; Harmon et al, 1999; Figure 1). Moreover, reduced MS activity due to MS c.2756A>G results in lower rates of synthesis of methionine and SAM, which might be protective against SAM-dependent DNA hypermethylation that is an additional risk factor for cancer development (Figure 1; Matsuo et al, 2001; Mudd et al, 2001): several studies have demonstrated an association of DNA hypermethylation of tumour-suppressor genes such as p53, p15 and p16 with lymphomas (Baur et al, 1999; Moller et al, 1999; Gonzalez et al, 2000).
The functional consequences of CBS 844ins68 are thought to be opposite to that of MS c.2756A>G. 844ins68 lowers CBS activity leading to an increase of homocysteine remethylation and reduced availability of 5,10-methylentetrahydrofolate for nucleic acid synthesis, while synthesis of SAM that serves as the substrate for DNA methylation is increased (Mudd et at, 2001; Figure 1). We found a higher frequency of CBS844ins68 in PCNSL patients than in controls. However, this difference did not reach statistical significance.
The MTHFR variants c.677C>T and c.1298A>C and their combinations have previously been reported to be associated with several types of human cancer (Ma et al, 1997; Skibola et al, 1999; Matsuo et al, 2001), but there were no significant associations of these polymorphisms with PCNSL in our study.
In summary, the results of this study suggest a protective function of MS c.2756A>G (G allele) against PCNSL and encourage future studies investigating the influence of genetic and dietary conditions of homocysteine and folate metabolism on cancerogenesis.
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Baur AS, Shaw P, Burri N, Delacretaz F, Bosman FT, Chaubert P (1999) Frequent methylation silencing of p15INK4b (MTS2) and p16INK4a (MTSl) in B-cell and T-cell lymphomas. Blood 94: 1773–1781
Blount BC, Mack MM, Wehr CM, MacGregor JT, Hiatt RA, Wang G, Wickramasinghe SN, Everson RB, Ames BN (1997) Folate deficiency causes uracil misincorporation into human DNA and chromosome breakage: implications for cancer and neuronal damage. Proc Natl Acad Sci USA 94: 3290–3295
Garter KC, Warnke RA (2001) Diffuse large cell lymphoma. In Pathology and Genetics of tumors of Haematopoietic and Lymphoid Tissues, Jaffe ES, Harris NL, Stein H (eds) pp 171–174. Lyon: IARC Press
Gonzalez M, Mateos MV, Garcia-Sanz R, Balanzategui A, Lopez-Perez R, Chillon MC, Gonzalez D, Alaejos I, San Miguel JF (2000) De novo methylation of tumor suppressor gene p16/INK4 is a frequent finding in multiple myeloma patients at diagnosis. Leukemia 14: 183–187
Gonzalez-Fraile MI, Garcia-Sanz R, Mateos MV, Balanzategui A, Gonzalez M, Vaquez L, Miguel JF (2002) Methylenetetrahydrofolate reductase genotype does not play a role in multiple myeloma pathogenesis. Br J Haematol 117: 890–892
Gonzalez-Gomez P, Bello MJ, Arjona D, Alonso ME, Lomas J, Aminoso C, de Campos JM, Sarasa JL, Gutierrez M, Rey JA (2003) CpG island methylation of tumour-related genes in three primary central nervous system lymphomas in immunocompetent patients. Cancer Genet Cytogenet 142: 21–24
Harmon DL, Shields DC, Woodside JV, McMaster D, Yarnell JW, Young IS, Peng K, Shane B, Evans AE, Whitehead AS (1999) Methionine synthase D919G polymorphism is a significant but modest determinant of circulating homocysteine concentrations. Genet Epidemiol 17: 298–309
Lincz LF, Scorgie FE, Kerridge I, Potts R, Spencer A, Enno A (2003) Methionine synthase genetic polymorphism MS A2756G alters susceptibility to follicular but not diffuse large B-cell non-Hodgkin's lymphoma or multiple myeloma. Br J Haematol 120: 1051–1054
Linnebank M, Homberger A, Junker R, Nowak-Goettl U, Harms E, Koch HG (2001) High prevalence of the I278T mutation of the human cystathionine β-synthase detected by a novel screening application. Thromb Haemost 85: 986–988
Linnebank M, Homberger A, Nowak-Gottl U, Marquardt T, Harms E, Koch HG (2000) Linkage disequilibrium of the common mutations 677C>T and 1298A>C of the human MTHFR gene as proven by the novel polymorphisms 129C>T, 1068C>T. Eur J Fed 159: 472–473
Ma J, Stampfer MJ, Christensen B, Giovannucci E, Hunter DJ, Chen J, Willett WC, Selhub J, Hennekens CH, Gravel R, Rozen R (1999) A polymorphism of the methionine synthase gene: association with plasma folate, vitamin B12, homocysteine, and colorectal cancer risk. Cancer Epidemiol Biomarkers Prev 8: 825–829
Ma J, Stampfer MJ, Giovannucci E, Artigas C, Hunter DJ, Fuchs C, Willet WC, Selhub J, Hennekens CH, Rozen R (1997) Methylenetetrahydrofolate reductase polymorphism, dietary interactions, and risk of colorectal cancer. Cancer Res 57: 1098–1102
Matsuo K, Suzuki R, Hamajima N, Ogura M, Kagami Y, Taji H, Kondoh E, Maeda S, Asakura S, Kaba S, Nakamura S, Seto M, Morishima Y, Tajima K (2001) Association between polymorphisms of folate- and methionine-metabolizing enzymes and susceptibility to malignant lymphoma. Blood 97: 3205–3209
Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16: 1215
Moller MB, Ino Y, Gerdes AM, Skjodt K, Louis DN, Pedersen NT (1999) Aberration of the p53 pathway components p53, MDM2 and CDKN2A appear independent in diffuse large B cell lymphoma. Leukemia 13: 453–459
Montesinos-Rongen M, Kuppers R, Schluter D, Spieker T, Van Roost D, Schaller C, Reifenberger G, Wiestler OD, Deckert-Schluter M (1999) Primary central nervous system lymphomas are derived from germinal-center B cells and show a preferential usage of the V4-34 gene segment. Am J Pathol 155: 2077–2086
Mudd SH, Levy HL, Kraus JP (2001) Disorders of transsulfuration. In The Metabolic and Molecular Bases of Inherited Disease, Scriver CR, Beaudet AL, Sly WS, Valle D, Childs B, Kinzler K, Vogelstein B (eds) pp 2007–2056. New York: McGraw-Hill
Nakamura M, Kishi M, Sakaki T, Hashimoto H, Nakase H, Shimada K, Ishida E, Konishi N (2003) Novel tumour suppressor loci on 6q22–23 in primary central nervous system lymphomas. Cancer Res 63: 737–741
Pels H, Schmidt-Wolf IGH, Glasmacher A, Schulz H, Engert A, Diehl V, Zellner A, Schackert G, Reichmann H, Kroschinsky F, Vogt-Schaden M, Egerer G, Bode U, Schaller C, Deckert M, Fimmers R, Helmstaedter C, Atasoyl A, Klockgether T, Schlegel U (2003) Primary CNS lymphoma: results of a pilot/phase II study of systemic and intraventricular chemotherapy with deferred radiotherapy. J Clin Oncol 21: 4489–4495
Skibola CF, Smith MT, Kane E, Roman E, Rollinson S, Cartwright RA, Morgan G (1999) Polymorphisms in the methylenetetrahydrofolate reductase gene are associated with susceptibility to acute leukemia in adults. Proc Natl Acad Sci USA 96: 12810–12815
Weber T, Weber RG, Kaulich K, Actor B, Meyer-Puttlitz B, Lampel S, Buschges R, Weigel R, Deckert-Schluter M, Schmiedek P, Reifenberger G, Lichter P (2000) Characteristic chromosomal imbalances in primary central nervous system lymphomas of the diffuse large B-cell type. Brain Pathol 10: 73–84
Acknowledgements
The technical assistance of Mihaela Keller is gratefully acknowledged. We thank Professor Dr Ruthild Weber, Department of Human Genetics, and Dr Bernd O Evert, Department of Neurology for critical reading of the manuscript.
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Linnebank, M., Schmidt, S., Kölsch, H. et al. The methionine synthase polymorphism D919G alters susceptibility to primary central nervous system lymphoma. Br J Cancer 90, 1969–1971 (2004). https://doi.org/10.1038/sj.bjc.6601777
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DOI: https://doi.org/10.1038/sj.bjc.6601777
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