Restricted accessResearch articleFirst published online 2012-01
Polymorphisms of Folate Pathway Enzymes (Methylenetetrahydrofolate Reductase and Thymidylate Synthase) and Their Relationship with Thymidylate Synthase Expression in Human Astrocytic Tumors
Two important polymorphisms of folate cycle enzymes, methylenetetrahydrofolate reductase (MTHFR) C677T and thymidylate synthase (TS) enhancer region (TSER) 28-bp tandem repeat, are related to risk of various types of cancer, including brain tumors, although there are few studies on this subject. A case–control study of these two polymorphisms in astrocytomas of different grades was carried out using polymerase chain reaction–restriction fragment length polymorphism, also determining the immunohistochemical expression of TS. The MTHFR 677 TT genotype was less associated with astrocytic tumors (odds ratio [OR]=0.00; p=0.0238), but the TSER polymorphism did not show any significant association. Combined genotype TT-double repeats/triple repeats (2R/3R) had a protective effect against astrocytomas (OR=0.00; p=0.0388). Expression of TS protein was observed in the majority of cases, with grade IV tumors being the exception. Moreover, the median H-score for the pilocytic astrocytomas was significantly higher when compared with that for diffuse tumors. There was an inverse correlation between the 2R/2R genotype and the highest TS-expressing tumors, and 3R/3R was relatively more frequent among the tumors grouped in the third and fourth quartiles. Our results provide support for the role of MTHFR and TS polymorphism in gliomagenesis, possibly because of the alteration of DNA methylation and repair status. Moreover, high levels of TS expression were detected in these tumors.
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References
1.
BethkeL., WebbE., MurrayA., SchoemakerM., FeychtingM., LönnS., AhlbomA., MalmerB., HenrikssonR., AuvinenA., KiuruA., SalminenT., JohansenC., ChristensenH.C., MuirK., McKinneyP., HepworthS., DimitropoulouP., LophatananonA., SwerdlowA., HoulstonR.2008. Functional polymorphisms in folate metabolism genes influence the risk of meningioma and glioma. Cancer Epidemiol Biomarkers Prev, 17:1195–1202.
2.
BlomekeB., BennettW.P., HarrisC.C., ShieldsP.G.1997. Serum, plasma and paraffinembedded tissues as sources of DNA for studying cancer susceptibility genes. Carcinogenesis, 18:1271–1275.
3.
BlountB.C., AmesB.N.1994. Analysis of uracil in DNA by gas chromatography-mass spectrometry. Anal Biochem, 219:195–200.
4.
BlountB.C., MackM.M., WehrC.M., Mac GregorJ.T., HiattR.A., WangG., WickramasingheS.N., EversonR.B., AmesB.N.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.
5.
BrandalizeA.P.C., BandinelliE., BorbaJ.B., FélixT.M., RoisenbergI., Schüler-FacciniL.2007. Polymorphisms in genes MTHFR, MTR and MTRR are not risk factors for cleft lip/palate in South Brazil. Braz J Med Biol Res, 40:787–791.
6.
CadieuxB., ChingT., VandenbergS.R., CostelloJ.F.2006. Genome-wide hypomethylation in human glioblastomas associated with specific copy number alteration, methylenetetrahydrofolate reductase allele status, and increased proliferation. Cancer Res, 1:8469–8473.
7.
CastroR., RiveraI., RavascoP., JakobsC., BlomH.J., CamiloM.E., de AlmeidaI.T.2003. 5,10-Methylenetetrahydrofolate reductase 677CT and 1298AC mutations are genetic determinants of elevated homocysteine. Q J Med, 96:297–303.
8.
ChenJ., HunterD.J., StampferM.J., KyteC., ChanW., WetmurJ.G., MosigR., SelhubJ., MaJ.2003. Polymorphism in the thymidylate synthase promoter enhancer region modifies the risk and survival of colorectal cancer. Cancer Epidemiol Biomarkers Prev, 12:958–962.
9.
ChungC.J., PuY.S., SuC.T., ChenH.W., HuangY.K., ShiueH.S., HsuehY.M.2010. Polymorphisms in one-carbon metabolism pathway genes, urinary arsenic profile, and urothelial carcinoma. Cancer Causes Control, 21:1605–1613.
10.
ClarkeJ., ButowskiN., ChangS.2010. Recent advances in therapy for glioblastoma. Arch Neurol, 67:279–283.
CuiL., ShinM.H., KweonS.S., KimH.N., SongH.R., PiaoJ.M., ChoiJ.S., ShimH.J., HwangJ.E., KimH.R., ParkY.K., KimS.H.2010. Methylenetetrahydrofolate reductase C677T polymorphism in patients with gastric and colorectal cancer in a Korean population. BMC Cancer, 10:2362–2368.
13.
DumitrescuR.G., CotarlaI.2005. Understanding breast cancer risk—where do we stand in 2005?J Cell Mol Med, 9:208–221.
14.
DuthieS.J.2010. Folate and cancer: how DNA damage, repair and methylation impact on colon carcinogenesis. J Inherit Metab Dis, 34:101–109.
15.
EtienneM-C., ChazalM., Laurent-PuigP., MagnéN., RostyC., FormentoJ.L., FrancoualM., FormentoP., RenéeN., ChamoreyE., BourgeonA., SeitzJ.F., DelperoJ.R., LetoublonC., PezetD., MilanoG.2002. Prognostic value of tumoral thymidylate synthase and p53 in metastatic colorectal cancer patients receiving fluorouracil-based chemotherapy: phenotypic and genotypic analyses. J Clin Oncol, 20:2832–2843.
FrosstP., BlomH.J., MilosR., GoyetteP., SheppardC.A., MatthewsR.G., BoersG.J.H., den HeijerM., KluijtmansL.A.J., van den HeuveL.P., RozenR.1995. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet, 10:111–113.
18.
Gonzalez-GomezP., BelloJ., ArjonaD., LomasJ., AlonsoM.E., De CamposJ.M., VaqueroJ., IslaA., GutierrezM., ReyJ.A.2003. Promoter hypermethylation of multiple genes in astrocytic gliomas. Int J Oncol, 22:601–608.
19.
GrazianoF., KawakamiK., WatanabeG., RuzzoA., HumarB., SantiniD., CatalanoV., FicarelliR., MerrimanT., PanunziS., TestaE., CascinuS., BearziI., ToniniG., MagnaniM.2004. Association of thymidylate synthase polymorphisms with gastric cancer susceptibility. Int J Cancer, 112:1010–1014.
20.
GrundaJ.M., FiveashJ., PalmerC.A., CantorA., Fathallah-ShaykhH.M., NaborsL.B., JohnsonM.R.2010. Rationally designed pharmacogenomic treatment using concurrent capecitabine and radiotherapy for glioblastoma; gene expression profiles associated with outcome. Clin Cancer Res, 16:2890–2898.
21.
GrundaJ.M., NaborsL.B., PalmerC.A., ChhiengD.C., StegA., MikkelsenT., DiasioR.B., ZhangK., AllisonD., GrizzleW.E., WangW., GillespieG.Y., JohnsonM.R.2006. Increased expression of thymidylate synthetase (TS), ubiquitin specific protease 10 (USP10) and survivin is associated with poor survival in glioblastoma multiforme (GBM)J Neurooncol, 80:261–274.
22.
HorieN., AibaH., OguroK., HojoH., TakeishiK.1995. Functional analysis and DNA polymorphism of the tandemly repeated sequences in the 5′-terminal regulatory region of the human gene for thymidylate synthase. Cell Struct Funct, 20:191–197.
23.
KafadarA.M., YilmazH., KafadarD., ErgenA., ZeybekU., BozkurtN., KudayC., IsbirT.2006. C677T gene polymorphism of methylenetetrahydrofolate reductase (MTHFR) in meningiomas and high-grade gliomas. Anticancer Res, 26:2445–2449.
24.
KawakamiK., WatanabeG.2003. Identification and functional analysis of single nucleotide polymorphism in the tandem repeat sequence of thymidylate synthase gene. Cancer Res., 63:6004–6007.
25.
KimH.N., LeeI.K., KimY.K., TranH.T., YangD.H., LeeJ.J., ShinM.H., ParkK.S., ShinM.G., ChoiJ.S., KimH.J.2008. Association between folate-metabolizing pathway polymorphism and non-Hodgkin lymphoma. Br J Haematol., 140:287–294.
26.
KimD.2007. The interactive effect of methyl-group diet and polymorphism of methylenetetrahydrofolate reductase on the risk of colorectal cancer. Mutat Res, 622:14–18.
27.
KimY.I.1999. Folate and carcinogenesis: evidence, mechanisms, and implications. J Nutr Biochem, 10:66–88.
28.
KristensenM.H., WeidingerM., BzorekM., PedersenP.L., MejerJ.2010. Correlation between thymidylate synthase gene variants, RNA and protein levels in primary colorectal adenocarcinomas. J Int Med Res, 38:484–497.
29.
LautenM., AsgedonG., WelteK., SchrappeM., StanullaM.2003. Thymidylate synthase gene polymorphism and its association with relapse in childhood B-cell precursor acute lymphoblastic leukemia. J Hematology, 88:353–354.
30.
LightfootT.J., SkibolaC.F., WillettE.V., SkibolaD.R., AllanJ.M., CoppedeF., AdamsonP.J., MorganG.J., RomanE., SmithM.T.2005. Risk of non–hodgkin lymphoma associated with polymorphisms in folate-metabolizing genes. Cancer Epidemiol Biomarkers Prev, 14:2999–3003.
31.
LiuY., SheteS., HoskingF.J., RobertsonL.B., BondyM.L., HoulstonR.S.2010. New insights into susceptibility to glioma. Arch Neurol, 67:275–278.
32.
MarshS., MallonM.A., GoodfellowP., McLeodH.L.2005. Concordance of pharmacogenetic markers in germline and colorectal tumor DNA. Pharmacogenomics, 6:873–877.
33.
MandolaM.V., StoehlmacherJ., Muller-WeeksS., CesaroneG., YuM.C., LenzH.J., LadnerR.D.2003. A novel single nucleotide polymorphism within the 5′ tandem repeat polymorphism of the thymidylate synthase gene abolishes USF-1 binding and alters transcriptional activity. Cancer Res., 63:2898–2904.
34.
McCartyK.S.Jr., SzaboE., FlowersJ.L., CoxE.B., LeightG.S., MillerL., KonrathJ., SoperJ.T., BudwitD.A., CreasmanW.T., SeigierH.F., McCartyK.S.Sr. 1986. Use of a monoclonal anti-estrogen receptor antibody in the immunohistochemical evaluation of human tumors. Cancer Res, 46:4244–4248.
35.
MeneiP., CapelleL., GuyotatJ., FuentesS., AssakerR., BatailleB., FrançoisP., Dorwling-CarterD., PaquisP., BauchetL., ParkerF., SabatierJ., FaisantN., BenoitJ.P.2005. Local and sustained delivery of 5-fluorouracil from biodegradable microspheres for the radiosensitization of malignant glioma: a randomized phase II trial. Neurosurgery, 56:242–248.
36.
PegoraroR.J., ChikosiA., RomL., RobertsC., MoodleyJ.2004. Methylenetetrahydrofolate reductase gene polymorphisms in black South Africans and the association with preeclampsia. Acta Obstet Gynecol Scand, 83:449–454.
37.
PonderB.A.J.2001. Cancer genetics. Nature, 411:336–341.
38.
ReidyJ.A.1987. Folate- and deoxyuridine-sensitive chromatid breakage may result from DNA repair during G2. Mutat Res, 192:217–219.
39.
SafarinejadM.R., ShafieiN., SafarinejadS.2010. Relationship between three polymorphisms of methylenetetrahydrofolate reductase (MTHFRC677T, A1298C, and G1793A) gene and risk of prostate cancer: a case-control study. Prostate, 70:1645–1657.
40.
SemmlerA., SimonM., MoskauS., LinnebankM.2006. The methionine synthase polymorphism c.2756A > G alters susceptibility to glioblastoma multiforme. Cancer Epidemiol Biomarkers Prev, 15:2314–2316.
41.
ShrubsoleM.J., GaoY.T., CaiQ., ShuX.O., DaiQ., HébertJ.R., JinF., ZhengW.2004. MTHFR polymorphisms, dietary folate intake, and breast cancer risk: results from the shanghai breast cancer study. Cancer Epidemiol Biomarkers Prev, 13:190–196.
42.
SirachainanN., WongruangsriS., KajanachumpolS., PakakasamaS., VisudtibhanA., NuchprayoonI., LusawatA., PhudhicharoenratS., ShuangshotiS., HongengS.2008. Folate pathway genetic polymorphisms and susceptibility of central nervous system tumors in Thai children. Cancer Detect Prev, 32:72–78.
43.
StevensV.L., McCulloughM.L., PavluckA.L., TalbotJ.T., FeigelsonH.S., ThunM.J., CalleE.E.2007. Association of polymorphisms in one-carbon metabolism genes and postmenopausal breast cancer incidence. Cancer Epidemiol Biomarkers Prev, 16:1140–1147.
44.
TanW., MiaoX., WangL., YuC., XiongP., LiangG., SunT., ZhouY., ZhangX., LiH., LinD.2005. Significant increase in risk of gastroesophageal cancer is associated with interaction between promoter polymorphisms in thymidylate synthase and serum folate status. Carcinogenesis, 26:1430–1435.
45.
UlrichC.M., CurtinK., PotterJ.D., BiglerJ., CaanB., SlatteryM.L.2005. Polymorphisms in the reduced folate carrier, thymidylate synthase, or methionine synthase and risk of colon cancer. Cancer Epidemiol Biomarkers Prev, 14:2509–2516.
46.
Van TriestB., PinedoH.M., GiacconeG., PetersG.J.2000. Downstream molecular determinants of response to 5-fluorouracil and antifolate thymidylate synthase inhibitors. Ann Oncol, 11:385–391.
47.
WajedA.S., LairdP.W., DeMeesterT.R.2001. DNA methylation: an alternative pathway to cancer. Ann Surg, 234:10–20.
48.
WangL., MiaoX., TanW., LuX., ZhaoP., ZhaoX., ShanY., LiH., LinD.2005. Genetic polymorphisms in methylenetetrahydrofolate reductase and thymidylate synthase and risk of pancreatic cancer. Clin Gastroenterol Hepatol, 3:743–751.
49.
WeberB.L., NathansonK.L.2000. Low penetrance genes associated with increased risk for breast cancer. Eur J Cancer, 36:1193–1199.
50.
WeissJ.R., BaerM.R., AmbrosoneC.B., BlancoJ.G., HutsonA., FordL.A., MoysichK.B.2007. Concordance of pharmacogenetic polymorphisms in tumor and germ line DNA in adult patients with acute myeloid leukemia. Cancer Epidemiol Biomarkers Prev, 16:1038–1041.
51.
WrenschM.R., BondyM.L., WienckeJ., YostM.1993. Environmental risk factors for primary malignant brain tumors: a review. J Neuro-oncol, 17:47–64.
52.
YangQ-H., BottoL.D., GallagherM., FriedmanJ.M., SandersC.L., KoontzD., NikolovaS., EricksonJ.D., SteinbergK.2008. Prevalence and effects of gene-gene and gene-nutrient interactions on serum folate and serum total homocysteine concentrations in the United States: findings from the third national health and nutrition examination survey DNA bank. Am J Clin Nutr, 88:232–246.
53.
YuK.H., WangW.X., DingY.M., LiH., WangZ.S.2008. Polymorphism of thymidylate synthase gene associated with its protein expression in human colon cancer. World J Gastroenterol, 14:617–621.
54.
YuanJ.M., LuS.C., Van Den BergD., GovindarajanS., ZhangZ.Q., MatoJ.M., YuM.C.2007. Genetic polymorphisms in the methylenetetrahydrofolate reductase and thymidylate synthase genes and risk of hepatocellular carcinoma. Hepatology, 46:749–758.
55.
ZhangZ., ShiQ., SturgisE.M., SpitzM.R., HongW.K., WeiQ.2004. Thymidylate synthase 5′- and 3′-untranslated region polymorphisms associated with risk and progression of squamous cell carcinoma of the head and neck. Clin Cancer Res, 10:7903–7910.
56.
ZhangZ., XuY., ZhouJ., WangX., WangL., HuX., GuoJ., WeiQ., ShenH.2005. Polymorphisms of thymidylate synthase in the 5′- and 3′-untranslated regions associated with risk of gastric cancer in South China: a case–control analysis. Carcinogenesis, 26:1764–1769.
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