High frequency of the 3R/3R polymorphism in the thymidylate synthase enhancer region in Indonesian childhood acute lymphoblastic leukemia
Keywords:
polymorphism, acute lymphoblastic leukemia, thymidilate synthase, methotrexate, children
Abstract
Background Deoxyuridylate monophosphate (dTMP) is neces-sary for DNA synthesis and thymidylate synthase (TS) is an im-
portant target of cancer chemotherapy. Ethnic variations of the
polymorphic tandem repeat sequence in the enhancer region of
the TS promoter has previously been described to influence the
outcome of acute lymphoblastic leukemia (ALL). A triple repeat is
associated with a higher TS gene expression than a double re-
peat, resulting in poorer outcome of ALL patients treated with anti-
folate methotrexate (MTX).
Objective In this study, we determined the incidences of TS and
methylenetetrahydrofolate reductase (MTHFR) polymorphism and
ethnic variations between Indonesian and Caucasian ALL cell
samples obtained at diagnosis. Furthermore, we determined the
involvement of TS polymorphisms in MTX sensitivity using a
thymidilate synthase inhibition assay (TSIA).
Methods ALL cell samples were obtained at diagnosis from 101
Indonesian and 157 Caucasian children treated with MTX prospec-
tively. Genotyping for TS and MTHFR was analyzed by Genescan
and Lightcycler. TS polymorphism was determined by PCR assay
and MTHFR polymorphism and was analyzed by melting curve
analyses on lightcycler.
Results Homozygous TS triple repeats were more than twice as
common in Indonesian samples (76.3%) than in Caucasian samples
(33.1%). Heterozygotes of the MTHFR mutations were seen in 15%
of the screened Indonesian samples.
Conclusion There are significant ethnic variations in TS gene
regulatory elements of leukemic cells. A difference was found be-
tween the MTX sensitivity and a double or triple repeat in the Cau-
casian ALL group. The samples with a triple repeat show a shift in
their distribution towards hypersensitivity to MTX. Further investi-
gation on Indonesian samples may give insight in the role of poly-
morphisms in MTX sensitivity
References
1. Rots MG, Willey JC, Jansen G, van Zontwijk CH,
Noordhuis P, De Muth Jp, et al. mRNA expression lev-
els of methotrexate resistancerelated proteins in child-
hood leukemia as determined by a standardized com-
petitive template based RT-PCR method. Leukemia
2000;14:2166-75.
2. Welsh SJ, Titley J, Brunton L, Valenti M, Monaghan P,
Jackman AL, et al. Comparison of thymidylate synthase
(TS) protein upregulation after exposure to TS inhibi-
tors in normal and tumor cell lines and tissues. Clin
Cancer Res 2000;6:2538-46.
3. Hori T, Ayusawa D, Shimizu K, Koyama H, Seno T.
Chromosome breakage induced by thymidylate stress
in thymidylate synthase-negative mutants of mouse
FM3A cells. Cancer Res 1984;44:703-9.
4. Hori T, Ayusawa D, Glover TW, Seno T. Expression of
fragile site on the human X chromosome in somatic
cell hybrids between human fragile X cells and
thymidylate synthase-negative mouse mutant cells. Jpn
J Cancer Res 1985;76:977-83.
5. Kaneda S, Takeishi K,Ayusawa D, Shimizu K, Seno T,
Altman S. Role in translation of a triple tandemly re-
peated sequence in the 5'-untranslated region of hu-
man thymidylate synthase mRNA. Nucleic Acids Res
1987;15;1259-70.
6. Horie N, Aiba H, Oguro K, Hojo H, Takeishi K. Func-
tional analysis and DNA polymorphism of the tandemly
repeated sequences in the 5'-terminal regulatory re-
gion of the human gene for thymidylate synthase. Cell
Struct Funct 1995;20:191-7.
7. Kawakami K, Omura K, Kanehira E, Watanabe Y. Poly-
morphic tandem repeats in the thymidylate synthase gene
is associated with its protein expression in human gas-
trointestinal cancers. Anticancer Res 1999;19:3249-52.
8. Pullarkat ST, Stoehlmacher J, Ghaderi V, Xiong YP,
Ingles SA, Sherrod A, et al. Thymidylate synthase gene
polymorphism determines response and toxicity of 5-
FU chemotherapy. Pharmacogenomics J 2001;1:65-70.
9. Johnston PG, Drake JC, Trepel J, Allegra CJ. Immuno-
logical quantitation of thymidylate synthase using the
monoclonal antibody thymidilate synthase 106 in 5-
fluorouracil-sensitive and resistant human cancer cell
lines. Cancer Res 1992;52,4306-12.
10. Marsh S, Collie-Duguid ES, Li T, Liu X, McLeod HL.
Ethnic variation in the thymidylate synthase enhancer
region polymorphism among Caucasian and Asian
populations. Genomics 1999;58:310-2.
11. Marsh S, Ameyaw MM, Githang’a J, Indalo A, Ofori-
Adjei D, Mcleod HL, et al. Novel thymidylate synthase
enhancer region alleles in African populations. Hum
Mutat 2000;16:528.
12. Krajinovic M, Costea I, Chiasson S. Polymorphism of
the thymidylate synthase gene and outcome of acute
lymphoblastic leukaemia. Lancet 2002;359:1033-4.
13. Hishida A, Matsuo K, Hamajima N, Ho H, Ogura M,
Kagami Y, et al. Associations between polymorphisms
in the thymidylate synthase and serine hydroxy-
methyltransferase genes and susceptibility to malignant
lymphoma. Haematologica 2003;88:159-66.
14. Skibola CF, Smith MT, Hubbard A Shane B, Robert
AC, Law GR, et al. Polymorphisms in the thymidylate
synthase and serine +hydroxymethyltransferase genes
and risk of adult acute lymphocytic leukemia. Blood
2002;99;3786-91.
15. Frosst P, Blan HJ, Milos R, Goyette P, Sheppard CA,
Mathews RG, et al. A candidate genetic risk factor for
vascular disease: A common mutation in methylene-
tetrahydrofolate reductase. Nat Genet 1995;10:111-3.
16. Scott J, Weir D. Folate/vitamin B12 inter-relationships.
Essays Biochem 1994;28:63-72.
17. Goyette P, Sumner JS, Milos R, Duncan AM,
Rosenblatt DS, Mathews RG, et al. Human methylene-
tetrahydrofolate reductase: Isolation of cDNA, map-
ping and mutation identification. Nat Genet 1994;7:
195-200.
18. Rozen R. Molecular genetics of methylenetetra-
hydrofolate reductase deficiency. J Inherit Metab Dis
1996;19:589-94.
19. Ueland PM, Hustad S, Schneede J, Refsum H, Vollset
SE. Biological and clinical implications of the MTHFR
C677T polymorphism. Trends Pharmacol Sci 2001;22:
195-201.
20. Bagley PJ, Selhub J. A common mutation in the
methylenetetrahydrofolate reductase gene is associ-
ated with an accumulation of formylated tetra-
hydrofolates in red blood cells. Proc Natl Acad Sci USA
1998;95:13217-20.
21. Kawakami K, Ishida Y, Danenberg KD, Omura,
Watanabe G, Danenberg PV, et al. Functional polymor-
phism of the thymidylate synthase gene in colorectal
cancer accompanied by frequent loss of heterozygos-
ity. Jpn J Cancer Res 2002;93:1221-9.
22. Rodenhuis S, Mcguire JJ, Sawicki WL, Bertino JR. Ef-
fects of methotrexate and of the nonclassical folate
antagonist trimetrexate on human-leukemia cells. Leu-
kemia 1987;1:116-20.
23. Rots MG, Pieters R, Kaspers HJ, van Zantwijk CH,
Noordhuis P, Mauritz R, et al. Differential methotrex-
ate resistance in childhood T-versus common/preB-
acute lymphoblastic leukemia can be measured by an
in situ thymidylate synthase inhibition assay, but not
by the MTT assay. Blood 1999;93:1067-74.
24. Etienne MC, Chazal M, Laurent-Puig P, Magne N,
Rosty C, Formento JL, et al. Prognostic value of tu-
moral thymidylate synthase and p53 in metastatic
colorectal cancer patients receiving fluorouracil-based
chemotherapy: Phenotypic and genotypic analyses. J
Clin Oncol 2002;20:2832-43.
25. Etienne MC, Ilc K, Formento JL, Laurent-Puig P,
Formento P, Cheradame S, et al. Thymidylate synthase
and methylenetetrahydrofolate reductase gene poly-
morphisms: relationships with 5-fluorouracil sensitiv-
ity. Br J Cancer 2004;90:526-34.
26. Lacopetta B, Grieu F, Joseph D, Elsaleh H. A polymor-
phism in the enhancer region of the thymidylate syn-
thase promoter influences the survival of colorectal
cancer patients treated with 5-fluorouracil. Br J Can-
cer 2001;85:827-30.
27. Mandola MV, Stoelmacher J, Muller-Weeks S, Cesarone
G, Yua MC, Lenz HJ, et al. A novel single nucleotide
polymorphism within the 5’tandem repeat polymor-
phism of the thymidylate synthase gene abolishes USF-
1 binding and alters transcriptional activity. Cancer
Res 2003;63:2898-904.
28. Ulrich CM, Yasui Y, Storb R, Schubert MM, Wagner
JL, Bigler J, et al. Pharmacogenetics of methotrexate:
Toxicity among marrow transplantation patients var-
ies with the methylenetetrahydrofolate reductase
C677T polymorphism. Blood 2002;98;231-4.
29. Urano W, Taniguchi A, Yamanaka H, Tanaka E,
Nakajima H, Matsuda Y, et al. Polymorphisms in the
methylenetetrahydrofolate reductase gene were asso-
ciated with both the efficacy and the toxicity of meth-
otrexate used for the treatment of rheumatoid arthri-
tis, as evidenced by single locus and haplotype analy-
ses. Pharmacogenetics 2002;12:183-90.
30. Sohn KJ, Croxford R, Yates Z, Lucock M, Kim YI. Ef-
fect of the methylenetetrahydrofolate reductase C677T
polymorphism on chemosensitivity of colon and breast
cancer cells to 5-fluorouracil and methotrexate. J Natl
Cancer Inst 2004;96:134-44.
31. Skibola CF, Smith MT, Kane E, Roman E, Rollinson
S, Cartwright RA, et al. Polymorphisms in the
methylenetetrahydrofolate reductase gene are asso-
ciated with susceptibility to acute leukemia in adults.
Proc Natl Acad Sci USA 1999;96;12810-5.
32. Wiemels JL, Smith RN, Taylor GM, Eden OB,
Alexander FE, Greaves MF. Methylenetetrahydrofolate
reductase (MTHFR) polymorphisms and risk of mo-
lecularly defined subtypes of childhood acute leuke-
mia. Proc Natl Acad Sci USA 2001;98:4004-9.
33. Krajinovic M, Lemieux-Blanchard E, Chiasson S,
Primeau M, Costea I, Moghrabi A. Role of polymor-
phisms in MTHFR and MTHFD1 genes in the out-
come of childhood acute lymphoblastic leukemia.
Pharmacogenomics J 2004;4:66-72.
Noordhuis P, De Muth Jp, et al. mRNA expression lev-
els of methotrexate resistancerelated proteins in child-
hood leukemia as determined by a standardized com-
petitive template based RT-PCR method. Leukemia
2000;14:2166-75.
2. Welsh SJ, Titley J, Brunton L, Valenti M, Monaghan P,
Jackman AL, et al. Comparison of thymidylate synthase
(TS) protein upregulation after exposure to TS inhibi-
tors in normal and tumor cell lines and tissues. Clin
Cancer Res 2000;6:2538-46.
3. Hori T, Ayusawa D, Shimizu K, Koyama H, Seno T.
Chromosome breakage induced by thymidylate stress
in thymidylate synthase-negative mutants of mouse
FM3A cells. Cancer Res 1984;44:703-9.
4. Hori T, Ayusawa D, Glover TW, Seno T. Expression of
fragile site on the human X chromosome in somatic
cell hybrids between human fragile X cells and
thymidylate synthase-negative mouse mutant cells. Jpn
J Cancer Res 1985;76:977-83.
5. Kaneda S, Takeishi K,Ayusawa D, Shimizu K, Seno T,
Altman S. Role in translation of a triple tandemly re-
peated sequence in the 5'-untranslated region of hu-
man thymidylate synthase mRNA. Nucleic Acids Res
1987;15;1259-70.
6. Horie N, Aiba H, Oguro K, Hojo H, Takeishi K. Func-
tional analysis and DNA polymorphism of the tandemly
repeated sequences in the 5'-terminal regulatory re-
gion of the human gene for thymidylate synthase. Cell
Struct Funct 1995;20:191-7.
7. Kawakami K, Omura K, Kanehira E, Watanabe Y. Poly-
morphic tandem repeats in the thymidylate synthase gene
is associated with its protein expression in human gas-
trointestinal cancers. Anticancer Res 1999;19:3249-52.
8. Pullarkat ST, Stoehlmacher J, Ghaderi V, Xiong YP,
Ingles SA, Sherrod A, et al. Thymidylate synthase gene
polymorphism determines response and toxicity of 5-
FU chemotherapy. Pharmacogenomics J 2001;1:65-70.
9. Johnston PG, Drake JC, Trepel J, Allegra CJ. Immuno-
logical quantitation of thymidylate synthase using the
monoclonal antibody thymidilate synthase 106 in 5-
fluorouracil-sensitive and resistant human cancer cell
lines. Cancer Res 1992;52,4306-12.
10. Marsh S, Collie-Duguid ES, Li T, Liu X, McLeod HL.
Ethnic variation in the thymidylate synthase enhancer
region polymorphism among Caucasian and Asian
populations. Genomics 1999;58:310-2.
11. Marsh S, Ameyaw MM, Githang’a J, Indalo A, Ofori-
Adjei D, Mcleod HL, et al. Novel thymidylate synthase
enhancer region alleles in African populations. Hum
Mutat 2000;16:528.
12. Krajinovic M, Costea I, Chiasson S. Polymorphism of
the thymidylate synthase gene and outcome of acute
lymphoblastic leukaemia. Lancet 2002;359:1033-4.
13. Hishida A, Matsuo K, Hamajima N, Ho H, Ogura M,
Kagami Y, et al. Associations between polymorphisms
in the thymidylate synthase and serine hydroxy-
methyltransferase genes and susceptibility to malignant
lymphoma. Haematologica 2003;88:159-66.
14. Skibola CF, Smith MT, Hubbard A Shane B, Robert
AC, Law GR, et al. Polymorphisms in the thymidylate
synthase and serine +hydroxymethyltransferase genes
and risk of adult acute lymphocytic leukemia. Blood
2002;99;3786-91.
15. Frosst P, Blan HJ, Milos R, Goyette P, Sheppard CA,
Mathews RG, et al. A candidate genetic risk factor for
vascular disease: A common mutation in methylene-
tetrahydrofolate reductase. Nat Genet 1995;10:111-3.
16. Scott J, Weir D. Folate/vitamin B12 inter-relationships.
Essays Biochem 1994;28:63-72.
17. Goyette P, Sumner JS, Milos R, Duncan AM,
Rosenblatt DS, Mathews RG, et al. Human methylene-
tetrahydrofolate reductase: Isolation of cDNA, map-
ping and mutation identification. Nat Genet 1994;7:
195-200.
18. Rozen R. Molecular genetics of methylenetetra-
hydrofolate reductase deficiency. J Inherit Metab Dis
1996;19:589-94.
19. Ueland PM, Hustad S, Schneede J, Refsum H, Vollset
SE. Biological and clinical implications of the MTHFR
C677T polymorphism. Trends Pharmacol Sci 2001;22:
195-201.
20. Bagley PJ, Selhub J. A common mutation in the
methylenetetrahydrofolate reductase gene is associ-
ated with an accumulation of formylated tetra-
hydrofolates in red blood cells. Proc Natl Acad Sci USA
1998;95:13217-20.
21. Kawakami K, Ishida Y, Danenberg KD, Omura,
Watanabe G, Danenberg PV, et al. Functional polymor-
phism of the thymidylate synthase gene in colorectal
cancer accompanied by frequent loss of heterozygos-
ity. Jpn J Cancer Res 2002;93:1221-9.
22. Rodenhuis S, Mcguire JJ, Sawicki WL, Bertino JR. Ef-
fects of methotrexate and of the nonclassical folate
antagonist trimetrexate on human-leukemia cells. Leu-
kemia 1987;1:116-20.
23. Rots MG, Pieters R, Kaspers HJ, van Zantwijk CH,
Noordhuis P, Mauritz R, et al. Differential methotrex-
ate resistance in childhood T-versus common/preB-
acute lymphoblastic leukemia can be measured by an
in situ thymidylate synthase inhibition assay, but not
by the MTT assay. Blood 1999;93:1067-74.
24. Etienne MC, Chazal M, Laurent-Puig P, Magne N,
Rosty C, Formento JL, et al. Prognostic value of tu-
moral thymidylate synthase and p53 in metastatic
colorectal cancer patients receiving fluorouracil-based
chemotherapy: Phenotypic and genotypic analyses. J
Clin Oncol 2002;20:2832-43.
25. Etienne MC, Ilc K, Formento JL, Laurent-Puig P,
Formento P, Cheradame S, et al. Thymidylate synthase
and methylenetetrahydrofolate reductase gene poly-
morphisms: relationships with 5-fluorouracil sensitiv-
ity. Br J Cancer 2004;90:526-34.
26. Lacopetta B, Grieu F, Joseph D, Elsaleh H. A polymor-
phism in the enhancer region of the thymidylate syn-
thase promoter influences the survival of colorectal
cancer patients treated with 5-fluorouracil. Br J Can-
cer 2001;85:827-30.
27. Mandola MV, Stoelmacher J, Muller-Weeks S, Cesarone
G, Yua MC, Lenz HJ, et al. A novel single nucleotide
polymorphism within the 5’tandem repeat polymor-
phism of the thymidylate synthase gene abolishes USF-
1 binding and alters transcriptional activity. Cancer
Res 2003;63:2898-904.
28. Ulrich CM, Yasui Y, Storb R, Schubert MM, Wagner
JL, Bigler J, et al. Pharmacogenetics of methotrexate:
Toxicity among marrow transplantation patients var-
ies with the methylenetetrahydrofolate reductase
C677T polymorphism. Blood 2002;98;231-4.
29. Urano W, Taniguchi A, Yamanaka H, Tanaka E,
Nakajima H, Matsuda Y, et al. Polymorphisms in the
methylenetetrahydrofolate reductase gene were asso-
ciated with both the efficacy and the toxicity of meth-
otrexate used for the treatment of rheumatoid arthri-
tis, as evidenced by single locus and haplotype analy-
ses. Pharmacogenetics 2002;12:183-90.
30. Sohn KJ, Croxford R, Yates Z, Lucock M, Kim YI. Ef-
fect of the methylenetetrahydrofolate reductase C677T
polymorphism on chemosensitivity of colon and breast
cancer cells to 5-fluorouracil and methotrexate. J Natl
Cancer Inst 2004;96:134-44.
31. Skibola CF, Smith MT, Kane E, Roman E, Rollinson
S, Cartwright RA, et al. Polymorphisms in the
methylenetetrahydrofolate reductase gene are asso-
ciated with susceptibility to acute leukemia in adults.
Proc Natl Acad Sci USA 1999;96;12810-5.
32. Wiemels JL, Smith RN, Taylor GM, Eden OB,
Alexander FE, Greaves MF. Methylenetetrahydrofolate
reductase (MTHFR) polymorphisms and risk of mo-
lecularly defined subtypes of childhood acute leuke-
mia. Proc Natl Acad Sci USA 2001;98:4004-9.
33. Krajinovic M, Lemieux-Blanchard E, Chiasson S,
Primeau M, Costea I, Moghrabi A. Role of polymor-
phisms in MTHFR and MTHFD1 genes in the out-
come of childhood acute lymphoblastic leukemia.
Pharmacogenomics J 2004;4:66-72.
Published
2016-10-18
How to Cite
1.
Ugrasena I, Sutaryo S, Supriadi E, Vroling L, Cloos J, Hooijberg J, Veerman A. High frequency of the 3R/3R polymorphism in the thymidylate synthase enhancer region in Indonesian childhood acute lymphoblastic leukemia. PI [Internet]. 18Oct.2016 [cited 26Apr.2024];46(3):103-2. Available from: https://paediatricaindonesiana.org/index.php/paediatrica-indonesiana/article/view/910
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Received 2016-10-13
Accepted 2016-10-13
Published 2016-10-18
Accepted 2016-10-13
Published 2016-10-18
