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MTHFD1 c.1958G>A and TCN2 c.776G>C polymorphisms of folate metabolism genes and their implication for oral cavity cancer

Katarzyna Malinowska
Katarzyna Malinowska
, 
Alicja Nowak-Zduńczyk
Alicja Nowak-Zduńczyk
, 
Anna Merecz-Sadowska
Anna Merecz-Sadowska
, 
Anna Szczepańska
Anna Szczepańska
, 
Dariusz Kaczmarczyk
Dariusz Kaczmarczyk
, 
Katarzyna Bliźniewska-Kowalska
Katarzyna Bliźniewska-Kowalska
 e 
Hanna Zielińska-Bliźniewska
Hanna Zielińska-Bliźniewska
   | 06 apr 2022
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Article Category: Orginal Study
Pubblicato online: 06 apr 2022
Pagine: 30 - 38
Ricevuto: 07 feb 2021
Accettato: 07 set 2021
DOI: https://doi.org/10.2478/ahem-2021-0041
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© 2022 Katarzyna Malinowska, et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1

Analysis of the genotype polymorphism MTHFD1 c.1958G>A after hydrolysis with restriction enzyme MspI; lane 1 - marker, lanes 3, 5, 7 - heterozygote GA, lanes 2, 4 - homozygous GG, lane 6 is homozygous AA
Analysis of the genotype polymorphism MTHFD1 c.1958G>A after hydrolysis with restriction enzyme MspI; lane 1 - marker, lanes 3, 5, 7 - heterozygote GA, lanes 2, 4 - homozygous GG, lane 6 is homozygous AA

Fig. 2

Analysis of genotype polymorphism TCN2 c.776G>C after hydrolysis with restriction enzyme ScrFI; lane 1 - marker; lanes 2, 3, 5, 6, 7 - heterozygous CG, lane 4 - homozygous CC, lane 8 is homozygous GG
Analysis of genotype polymorphism TCN2 c.776G>C after hydrolysis with restriction enzyme ScrFI; lane 1 - marker; lanes 2, 3, 5, 6, 7 - heterozygous CG, lane 4 - homozygous CC, lane 8 is homozygous GG

Relationships between the patient group and the control group regarding smoking, alcohol consumption, and gender

Variables value Controls (%) Patients (%)N=220/439 OR (95%CI) P value
Tobacco
(N=220)
Non-smokers 150 (75.0) 31 (14.1) Reference
Smokers 50 (25.0) 189 (85.9) 18.29 (11.13–30.06) <0.0001
(N=439)
Non-smokers 150 (75.0) 124 (28.2) Reference
Smokers 50 (25.0) 315 (71.8) 7.62 (5.20–11.16) <0.0001
Alcohol consumption
(N=220)
No 101 (50.5) 70 (31.8) Reference
Yes 99 (49.5) 150 (68.2) 2.19 (1.47–3.25) <0.0001
(N=439)
No 101 (50.5) 149 (33.9) Reference
Yes 99 (49.5) 290 (66.1) 1.98 (1.41–2.79) <0.0001
Gender
(N=220)
Female 100 (50.0) 52 (23.6) Reference
Male 100 (50.0) 168 (76.4) 3.23 (2.13–4.90) <0.0001
(N=439)
Female 100 (50.0) 124 (28.2) Reference
Male 100 (50.0) 315 (71.8) 2.54 (1.80–3.59) <0.0001

Genotype and allele frequencies of MTHFD1 c.1958G>A and TCN2 c.776G>C polymorphisms in oral cavity cancer patients and controls

Genotypes Controls (%)N=200 Patients (%)N=220/439 OR (95% CI) P value
MTHFD1c.1958G>A
GG 53 (26.5) 55 (12.5) 1.00
GA 110 (55) 332 (75.6) 2.91 (1.88–4.49) <0.0001
AA 37 (18.5) 52 (11.9) 1.35 (0.77–2.38) 0.29
G 216 (54) 442 (49) 1.00
A 184 (46) 436 (51) 1.16 (0.91–1.47) 0.23
TCN2 c.776G>C
GG 34 (17) 76 (34.5) 1.00
CG 71 (35.5) 107 (48.7) 0.67 (0.41–1.12) 0.13
CC 95 (47.5) 37 (16.8) 0.17 (0.10–0.30) <0.0001
G 139 (35) 259 (59) 1.00
C 261 (65) 181 (41) 0.76 (0.58–1.01) 0.06

Detailed characterization of MTHFD1 and TCN2 gene polymorphisms

Gene The encoded protein Number of polymorphism Location Function of polymorphism1 Allele MAF
MTHFD1 Methylene-tetrahydrofolate-dehydrogenase rs2236225 14: 63978598 Arg653Gln G/A 0,46
TCN2 Transcobalamin 2 rs1801198 22: 29341610 Arg259Pro G/C 0,45

The primers, length of PCR products, and restriction enzymes

Polymorphism Primers Product length Restriction enzyme
MTHFD1 c.1958G>A Sense5′-CAC TCC AGT GTT TGT CCA TG-3′Antisense5′-GCA TCT TGA GAG CCC TGA C-3′ 331 bp MspI(37°C)
TCN2 c.776G>C Sense5′-CAC TTC AAG ACA CAG CAG CCA-3′Antisense5′-TCC CCT CAC TGG GAC GTC AAT-3′ 568 bp ScrFI(37°C)

Odds ratio of oral cavity cancer related to MTHFD1 and TCN2 genotypes according to tobacco use, alcohol consumption and gender in patients with head and neck cancer

Risk factor OR (95%CI) - P value
MTHFD1c.1958G>A * TCN2 c.776G>C **
Tobacco
Non-smokers 2.10 (1.09–4.06) P=0.03 0.72 (0.29–1.77) P=0.63
Smokers 2.60 (1.29–5.23) P=0.01 0.15 (0.05–0.45) P<0.0001
Alcohol consumption
No 1.23 (0.60–2.51) P=0.35 0.53 (0.27–1.01) P=0.08
Yes 1.01 (0.57–1.81) P=1 0.26 (0.13–0.52) P<0.0001
Gender
Female 1.77 (0.93–3.39) P=0.1 0.48 (0.22–1.04) P=0.07
Male 2.28 (1.32–3.96) P=0.003 0.33 (0.17–0.61) P=0.003
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Life Sciences, Molecular Biology, Microbiology and Virology, Medicine, Basic Medical Science, Immunology
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