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

Katarzyna Malinowska; Alicja Nowak-Zduńczyk; Anna Merecz-Sadowska; Anna Szczepańska; Dariusz Kaczmarczyk; Katarzyna Bliźniewska-Kowalska; Hanna Zielińska-Bliźniewska

Dziennik i szczegóły wydania

Postępy Higieny i Medycyny Doświadczalnej

Informacje o czasopiśmie

Format: Czasopismo
eISSN: 1732-2693
Pierwsze wydanie: 20 Dec 2021
Częstotliwość wydawania: 1 raz w roku
Języki: Angielski

Cancers of the head and neck can damage the brain, spinal cord, and nerves, as well as the sense organs responsible for contact with the outside world. Oncogenic transformation occurs following mutations that change the function of specific genes, such tumor suppressor genes or oncogenes, and their encoded protein products. Therefore, the aim of this study was to evaluate the relationship between the occurrence of MTHFD1 c.1958G>A and TCN2 c.776G>C gene polymorphisms and the risk of oral cavity cancer.

The study population consisted of 439 patients and 200 healthy subjects. Genomic DNA was extracted from peripheral blood and from paraffin-embedded tissue. Analysis of the gene polymorphisms was performed using polymerase chain reaction–restriction fragment length polymorphism.

Statistically significant differences were found in the distribution of genotypes of the rs2236225 and rs1801198 polymorphisms between patients and controls. Regarding MTHFD1 c.1958G>A, the GA genotype (p<0.0001, OR = 2.91, 95% CI = 1.88–4.49-GA) was more common among patients than healthy subjects. Regarding TCN2 c.776G>C, the frequency of CC genotype (p< 0.0001, OR = 0.17, 95% CI = 0.10–0.33-CC) was significantly less common among patients than healthy subjects. Tobacco smoking, alcohol use, and male gender (p<0.0001) were found to be predictors of the disease.

The results suggest that the MTHFD1 c.1958G>A polymorphism may be associated with a greater risk of oral cavity cancer, whereas a protective effect has been found for the TCN2 c.776G>C polymorphism.

Keywords

polymorphism
gene
gene
oral cavity cancer

Artykuł

Introduction

Head and neck cancer is the sixth most commonly diagnosed cancer, making it a serious and growing health problem [1]. The term head and neck cancer encompasses a very broad group of cancers caused by tumor growth in parts of the upper respiratory tract, such as the nasal cavity, paranasal sinuses, oral cavity, pharynx, and larynx [2]. The symptoms of head and neck cancer differ depending on the location of origin of the cancer. Common symptoms are hoarseness, ulceration, pain, and weight loss for cancers of the larynx, oral cavity, oropharynx, and hypopharynx, respectively [3]. The cancers are classified within the same group due to the similarity of the diagnostic and therapeutic tools used for their detection; they are classified according to the anatomically based TNM classification of malignant tumors [4]. More than 90% of head and neck cancers are squamous cell carcinoma (HNSCC) [1]. Other malignant neoplasms are diagnosed much less frequently, most often they are salivary gland tumors or even more rarely soft tissue tumors or lymphomas.

In 2018, tumors of the lip and oral cavity accounted for 2% of all malignant tumors registered worldwide (354,864 cases) and contributed to 177,384 deaths that year [4]. Epidemiological evidence indicates that the worldwide incidence of head and neck cancer increases with age, occurring most often in people over 50 years old, and they are more common in men than women. The risk of cancer is caused mainly by exposure to tobacco and alcohol abuse [1, 5]. Infection with human papilloma virus [6], environmental factors [7], dietary habits [8], and poor oral hygiene [9] also play a role in the pathogenesis of head and neck cancer. Intensive worldwide, multicenter research has been undertaken to discover new prognostic and treatment predictive biomarkers for these cancers [10, 11, 12, 13].

Carcinogenesis is a complex and multistage process which involves the conversion of normal cells into tumor cells. Oncogenic transformation occurs following mutations that change the function of specific genes, such tumor suppressor genes or oncogenes, and their encoded protein products. Important changes in the transcriptional and translational regulation of genes also occur in the process of malignant transformation [14]. Recently, much attention has been given to epigenetic changes that influence the regulation of gene expression without affecting the sequence of nucleotides. These phenomena are reversible under the action of chemicals and act through two routes: the epigenetic modification of gene expression by DNA methylation and the remodeling of chromatin by the alteration of histone proteins [15]. DNA methylation is essential for normal cell growth and differentiation. That process occurs during X-chromosome inactivation, genomic imprinting, cell death, and cell cycle control. In mammalian cells, cytosine residues of CpG sites are methylated by methyltransferase (DNMT) at position C5 [16]. The methylation process plays an important role in genome stability. Disorders of methylation patterns may become useful biological markers associated with many forms of tumor. Therefore, despite being difficult to clearly define and with no well-defined mechanism of action, epigenetic phenomena are of great potential clinical importance [17].

DNA methylation is a biochemical process of adding a methyl group to the substrate. The following enzymes take part in that process: methylenetetrahydrofolate reductase (MTHFR) [18], methylenetetrahydrofolate dehydrogenase, cyclohydrolase and formyltetrahydrofolate synthetase 1 (MTHFD1) [19], methionine synthase (MTR) [20], methionine synthase reductase (MTRR) [21], transcobalamin 2 (TCN2) [22], methyltransferase betaine – homocysteine (BHMT) [23], methyltransferase betaine – homocysteine 2 (BHMT2) [24] and synthase β – cystathionine (CBS) [25]. Numerous studies implicate abnormal methylation in cancer. Aberrant process may be related to the occurrence of a polymorphism in the gene encoding the corresponding protein.

MTHFD1 is an enzyme that performs three different activities (5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrolase, and 10-formylotetrahydrofolate synthetase). Each of these catalyzes the interconversion of tetrahydrofolate. That is a substrate for the biosynthesis of methionine, thymidylate, and purine nucleotides. The most analyzed polymorphism of the MTHFD1 gene, c.1958G>A, is located within the domain responsible for 10-formyltetrahydrofolate synthetase activity. The polymorphism results in the substitution of glutamine (Gln) with arginine (Arg) at codon 653 [19].

TCN2 belongs to the vitamin B₁₂ – binding protein family and allows cobalamin to be transported into the cell [26]. Vitamin B₁₂ is a co-factor for the enzyme MTR, which catalyzes the transfer of a methyl group from 5-methyltetrahydrofolate. That chemical as well as methionine are converted by MTHFR from 5,10-methylenetetrahydrofolate. Methionine may be then converted to S-adenosylmethionine, an important methyl donor for the methylation process, including DNA methylation. MTR links the methylation and synthesis of purine and pyrimidine pathways via generation of the methyl donor methionine and synthesis of tetrahydrofolate, respectively [27]. Both low MTR activity and low vitamin B₁₂ level are determinants of DNA hypomethylation [28]. The most common TCN2 polymorphism is the replacement of guanine to cytosine at position 776; this change will replace the amino acid arginine (Arg) with proline (Pro) at position 259 in the protein sequence of TCN2. This change in the TCN2 protein sequence affects the ability of the protein to bind to vitamin B₁₂ and disrupts the binding of the transcobalamin-vitamin B₁₂ receptor complex. The TCN2 c.776G>C genotype has a significant influence on the cellular delivery system of vitamin B₁₂ [29]. Healthy subjects with low vitamin B₁₂ status manifest reduced levels of homocysteine [30].

Our previous study [31] showed that the TCN2 gene polymorphism modulates the risk of oral cancer, but the number of patients and controls were limited. Recent studies have examined whether polymorphisms within the MTHFD1 and TCN2 genes correlate with increased risk of oral cavity cancer and may be associated with cancer risk, tobacco smoking, alcohol consumption and gender.

Materials and Methods

1.1

Patients

The study was conducted with the approval of the Local Ethics Committee of the Medical University of Lodz (RNN/142/09/KB). A total of 439 unrelated patients (201 women and 238 men) with histologically confirmed oral cavity squamous cell carcinoma (OCSCC) were examined for the MTHFD1 c.1958G>A polymorphism and 220 patients (97 women and 123 men) for the TCN2 c.776G>C polymorphism. The control group consisted of 200 (100 women and 100 men) unrelated patients with no confirmed malignancies, being treated for such diseases as nasal septum deviation, palatine uvula hypertrophy with sleep apnea, or epistaxis. Four hundred and fifteen (64.9%) participants were men (315 patients and 100 controls) and two hundred and twenty-four (35.1%) were women (124 patients and 100 controls). Alcohol was consumed by 68% of the experimental group, compared to 15% of the controls. Tobacco use also differed greatly between cases: 72% of the experimental group and 25% of the control group. The age of the patients in the study group ranged from 42 to 90 years (average age 63.7 years), and the control group was aged 26 to 84 years (average age 52.4 years). All subjects were cross-matched regarding age and sex. Samples of peripheral blood (5 ml) were taken in EDTA from the antecubital vein. A sample of postoperative tumor tissue was also taken from the mouth of all subjects in the experimental group and parts of the samples were secured in paraffin blocks. All study materials were collected in the Department of Otolaryngology, Medical University of Lodz, Poland. Informed, written consent to participate in the study was obtained from all patients.

1.2

Polymorphism analysis

MTHFD1 c.1958G>A genotyping by PCR-RFLP

DNA was extracted from peripheral blood lymphocytes using Genomic Mini AX Blood Kits (A&A Biotechnology, Gdynia, Poland) and from tissues in the paraffin blocks using Genomic Mini AX Tissue (A&A Biotechnology, Gdynia, Poland). Genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). PCR was carried out in a volume of 10 µl. The reaction mixture consisted of 100 ng of genomic DNA, 0.5 µmol of each primer, and 3U Taq polymerase (A&A Biotechnology, Gdynia, Poland). The PCR cycling conditions consisted of an initial denaturation step of 95°C for 5 minutes, followed by 35 cycles of 95°C for 30 s, 62°C for 30 s, 72°C for 30 s and then a final extension step at 72°C for 5 minutes (Bio-Rad, Hercules, California, USA). Following this, 10 µl of specifically positive PCR products was digested overnight with 1 µl of MspI (New England Bio Labs, Ipswich, Massachusetts, USA) at 37°C, and the digested DNA fragments were resolved on a 3% agarose gel (Sigma-Aldrich, Saint Louis, Missouri, USA). Obtained restrictive fragments were visualized by staining with ethidium bromide (Sigma-Aldrich, Saint Louis, Missouri, USA) and imaged under a UV light source (Bio-Rad, Hercules, California, United States). Genotypes were designated as follows: 197, 70, 56, and 8 bp for G/G; 267, 197, 70, 56, 8 bp for G/A; and 267 bp for A/A. Figure 1 presents a representative gel obtained after digestion of PCR.

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

1.3

TCN2c.776G>C genotyping by PCR-RFLP

DNA was extracted from peripheral blood lymphocytes using DNA Blood Mini Kits (A&A Biotechnology, Gdynia, Poland). Genotypes were determined by PCR-RFLP. PCR was carried out in a volume of 10 µl. The reaction mixture consisted of 100 ng of genomic DNA, 0.5 µmol of each primer, and 3 U Taq polymerase (A&A Biotechnology, Gdynia, Poland). The PCR was conducted under the following temperature conditions: pre-denaturation step at 94°C for 10 min, 34 cycles of denaturation at 56 °C for 45s oligonucleotide annealing at 55 °C for 30 s, and amplification at 72°C for 45 s, and final incubation at 72 °C for 10 min (Bio-Rad, Hercules, California, USA). Subsequently, the products were subjected to digestion by the ScrFI restriction enzyme (New England Bio Labs, Ipswich, Massachusetts, USA) at 37 °C. After 16 h of digestion, the samples were separated by electrophoresis (Bio-Rad, Hercules, California, USA) on a 2% agarose gel (Sigma-Aldrich, Saint Louis, Missouri, USA). Obtained restrictive fragments were visualized by staining with ethidium bromide (Sigma-Aldrich, Saint Louis, Missouri, USA) and imaged under a UV light source (Bio-Rad, Hercules, California, United States). Genotypes were designated as follows: 204 bp and 364 bp for C/C; 204, 364 and 568 bp for C/G; and 568 bp for G/G. Figure 2 presents a representative gel obtained after digestion of PCR products.

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

The primers, lengths of the PCR products, and restriction enzymes of MTHFD1 and TCN2gene polymorphisms are summarized in Table 1. Detailed characterization has been shown in Table 2. The MTHFD1 c.1958G>A and TCN2 c.776G>C polymorphisms were investigated according to Cheng et al. [32] and Pietrzyk et al. [33] with some alterations.

Table 1

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)

MTHFD1 - methylenetetrahydrofolate dehydrogenase, cyclohydrolase, and formyltetrahydrofolate synthetase 1; TCN2 - transcobalamin 2

Table 2

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

According to the Single Nucleotide Polymorphism Database (dbSNP), NCBI

MAF - minor allele frequency; MTHFD1 - methylenetetrahydrofolate dehydrogenase, cyclohydrolase, and formyltetrahydrofolate synthetase 1; TCN2 - transcobalamin 2

Statistical analysis

Multiple logistic regression was used to determine the effect of the interaction between genetic polymorphisms and risk factors associated with head and neck cancer. The model includes gender (reference: female), the consumption of tobacco products (reference: nonsmoking), and alcohol consumption (reference: nondrinkers). The frequencies of the analyzed polymorphisms among OCSCC patients and controls were evaluated using the Hardy-Weinberg (HW) equilibrium test. The distribution of the alleles was analyzed with the Yates chi-square test. Results are shown as odds ratio (OR) and 95% confidence intervals (95% CI). P-values were calculated as two-sided. Means were compared using a t-test or analysis of variance. Probabilities were considered significant at p values less than 0.05 (Statistica, Tulsa, Oklahoma, USA).

Results

The genotype and allele distributions of the MTHFD1 c.1958G>A and TCN2 c.776G>C gene polymorphisms are summarized in Table 3. The observed genotype frequencies of MTHFD1 c.1958G>A and TCN2 c.776G>C in the control group were in agreement with the Hardy-Weinberg equilibrium (HWE).

Table 3

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

OR - odds ratio; N – number; P value - probability value; MTHFD1 - methylenetetrahydrofolate dehydrogenase, cyclohydrolase, and formyltetrahydrofolate synthetase 1; TCN2 - transcobalamin 2

For the MTHFD1 c.1958G>A polymorphism, GG, GA, and AA genotype frequencies were 12.5, 75.6, and 11.9%, respectively, for the subjects who had suffered from cancer and 26.5, 55 and 18.5%, respectively, for control patients. The 1958 G allele frequencies were 49% among the subjects who had suffered from cancer and 54% among the control patients, while the 1958 A allele frequencies were 51% among subjects who had suffered from cancer and 46% among controls.

For the TCN2 c.776G>C polymorphism, GG, CG, and CC genotype frequencies were 34.5%, 48.7%, and 16.8% for the experimental group, and 17%, 35.5%, and 47.5%, respectively, for the control. In the gene TCN2, 776G allele frequencies were 59% among the subjects who had suffered from cancer and 35% among the controls, while 776C allele frequencies were 41% among subjects who had suffered from cancer and 65% among controls.

The results of comparison between groups showed that tobacco, alcohol use, and male gender (p < 0.0001) were predictors of the disease (Table 4).

Table 4

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

OR - odds ratio; N- number; P value - probability value

The data of the interaction between genotype and tobacco smoking, alcohol consumption, and gender with respect to the risk for oral cavity cancer are shown in Table 5. The presence of the MTHFD1 c.1958G>A variant genotype may increase the risk of oral cavity cancer for smokers (p= 0.01) and males (p= 0.003). On the other hand, the protective effect was observed for TCN2 c.776G>C variant genotype for smokers (p < 0.0001), alcohol consumption (p< 0.0001), and males (p = 0.003).

Table 5

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

OR - odds ratio; P value - probability value; MTHFD1 - methylenetetrahydrofolate dehydrogenase, cyclohydrolase and formyltetrahydrofolate synthetase 1; TCN2 - transcobalamin 2

MTHFD1c.1958G>A Reference: GG wild type genotype vs. variants GA+GG

TCN2 c.776G>C Reference: GG wild type genotype vs. variants GC+CC

Discussion

Malignant tumors have a significant impact on public health and constitute a significant economic burden on the population due to lost productivity and the costs associated with illness and treatment. It is expected that this problem will grow as the population ages [34].

He ad and neck cancers are classified according to their location. Risk factors, diagnosis, and treatment may vary depending on the disease subtype. The head and neck region of the body is located close to other important life organs and systems. Cancers of the head and neck can damage the brain, spinal cord, and nerves, as well as the sense organs responsible for contact with the outside world. The tumors may also affect the endocrine organs, such as the thyroid or pituitary gland. Regardless of whether a tumor is malignant or benign, it often directly affects the performance of several structures simultaneously [2].

Non-healing ulcers, new or fast-growing growths, or warts on the skin, mouth, or nose can develop in response to anxiety and need to be investigated to determine whether they pose a threat to health or life. Particular attention should be paid to frequently occurring headaches, dizziness and changes in temperament and mood. In addition, frequent nosebleeds, ear or neck aches, persistent hoarseness, and voice alterations can be symptoms of cancer, as can problems with chewing and swallowing, choking, and the presence of ulcers in the mouth and blood in the saliva. Other symptoms may include numbness in the face or tongue, tooth loss, or a well-fitting prosthesis becoming mismatched [35].

Al though genetic and environmental factors take part in the pathogenesis of head and neck cancer, the mechanisms underlying the process remain unclear. However, host genetic factors are the important determinants of the pathophysiology of the disease. Polymorphisms may be potentially used as a biomarker to evaluate cancer risk. Sequence variants in Toll-like receptor genes, which mediate the innate immune response defense against microorganisms, have been linked to head and neck cancer risk [36]. Moreover, strong evidence confirms that alterations in the genes associated with DNA methylation may contribute to carcinogenesis. Hypermethylation promotes gene silencing, and hypomethylation of a promoter can make it susceptible to induction by oncogenes [37].

The aim of the study was to determine the relationship between the occurrence of two polymorphisms of genes encoding selected enzymes associated with the methylation process, viz. MTHFD1 (rs2236225) and TCN2 (rs1801198), and the risk of developing cancer of the head and neck. This study included a group of 439 patients with histologically confirmed oral cavity cancer and a control group of 200 individuals without cancer.

Data indicate the presence of 30,000 CpG dinucleotides, also known as CpG islands, in the human genome; most of which are located in promotor regions. Under normal conditions, they remain unmethylated. Methylation of those structures leads to different diseases, including carcinogenesis [38]. Hazra et al. [39] found that aberrant methylation of the promoter CpG island may contribute to colorectal cancer development by silencing tumor suppressor genes. The inactivation of tumor suppressor genes by hypermethylation plays a key role in the process of malignant transformation [40].

MTHFR c. 677C>T gene polymorphism may be a risk factor for thyroid cancer. Furthermore, MTR c. 2756A>G is associated with tumor extent and aggressiveness [41]. Moreover, DNMT3B c. -283T>C gene polymorphism is linked to a higher risk of head and neck cancer [42]. The available literature data demonstrates the impact of the MTHFD1 c.1958G>A gene polymorphism on the appearance of neural tube birth defects in children [43, 44] and loss in the second trimester of pregnancy [45]. Data indicate that SNPs in the MTHFD gene coexist with folate and homocysteine concentration disorders in families with congenital heart disease [46]. Kempisty et al. reported a higher prevalence of schizophrenia and bipolar disorder among patients with 1958AA and 1958AG genotypes [47], whereas Sutherland et al. found no evidence for an association of the MTHFD1 c.1958G>A polymorphism with migraine susceptibility [48]. The MTHFD1 gene polymorphism is also regarded as the most common risk factor for cancer. Stevens et al. [49] noted an increased risk of developing malignant breast cancer in postmenopausal women who were carriers of the T and C alleles in the presence of at least one variant of the MTHFD1 c.677C>T and MTHFD1 c.1298A>C, respectively. Moreover, breast cancer patients who were carrying A allele for the MTHFD1 c.1958G>A polymorphism showed a higher frequency of tumor CpG island hypermethylation [50]. Wang et al. reported a connection between the presence of several mutated variants of this gene and the increased risk of gastric cancer [51]. Another study found the MTHFD1 c.1958G>A polymorphism to be related with a lower probability of event-free survival in children with lymphoblastic leukemia [52]. Despite expectations, MTHFD1c.1958G>A polymorphism has been found not to be a risk factor for prostate cancer [53]; however, MTHFD1 c.1958G>A SNP was associated with the emergence of squamous cancer cells in the head and neck in alcohol abusers and smokers [54].

It has been found that the presence of the TCN2 c.776CG+GG variant was related to a higher risk of occurrence of other disease entities such as colorectal adenoma [55] and Crohn disease [56]. The TCN2 776GG variant increased the susceptibility of individuals to peripheral nerve dysfunction [57]. Our earlier results indicate that the CG and GG genotypes and the G allele increase the likelihood of oral cancer [31]. Protein encoded by the TCN2c.776G>C G allele is associated with lower vitamin B₁₂ transport from blood to tissues [30], and patients with GG genotype had significantly lower concentrations of holotranscobalamin [58].

In conclusion, the presence of the MTHFD1 c.1958G>A polymorphism increases the risk of head and neck cancer associated with tobacco smoking and male sex, whereas TCN2 c.776G>C variant has protective effects with tobacco smoking, alcohol consumption, and male sex.

Ilustracje i tabele

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

Referencje

[1] Vigneswaran N, Williams MD. Epidemiologic trends in head and neck cancer and aids in diagnosis. Oral Maxillofac. Surg. Clin. North Am. 2014; 26: 123–141. VigneswaranN WilliamsMD Epidemiologic trends in head and neck cancer and aids in diagnosis Oral Maxillofac. Surg. Clin. North Am. 2014 26 123 141 10.1016/j.coms.2014.01.001404023624794262 Search in Google Scholar

[2] Heroiu Cataloiu AD, Danciu CE, Popescu CR. Multiple cancers of the head and neck. Maedica. 2013; 8: 80–85. Heroiu CataloiuAD DanciuCE PopescuCR Multiple cancers of the head and neck Maedica. 2013 8 80 85 Search in Google Scholar

[3] Douglas CM, Ingarfield K, McMahon AD, Savage SA, Conway DI, MacKenzie K. Presenting symptoms and long-term survival in head and neck cancer. Clin. Otolaryngol. 2018; 43: 795–804. DouglasCM IngarfieldK McMahonAD SavageSA ConwayDI MacKenzieK Presenting symptoms and long-term survival in head and neck cancer Clin. Otolaryngol. 2018 43 795 804 10.1111/coa.1305329247495 Search in Google Scholar

[4] Huang SH, O’Sullivan B. Overview of the 8th edition TNM classification for head and neck cancer. Curr. Treat. Options Oncol. 2017; 18: 40. HuangSH O’SullivanB Overview of the 8th edition TNM classification for head and neck cancer Curr. Treat. Options Oncol. 2017 18 40 10.1007/s11864-017-0484-y28555375 Search in Google Scholar

[5] Beynon RA, Lang S, Schimansky S, Penfold CM, Waylen A, Thomas SJ, Pawlita M, Waterboer T, Martin RM, May M, et al. Tobacco smoking and alcohol drinking at diagnosis of head and neck cancer and all-cause mortality: Results from head and neck 5000, a prospective observational cohort of people with head and neck cancer. Int. J. Cancer. 2018; 143: 1114–1127. BeynonRA LangS SchimanskyS PenfoldCM WaylenA ThomasSJ PawlitaM WaterboerT MartinRM MayM Tobacco smoking and alcohol drinking at diagnosis of head and neck cancer and all-cause mortality: Results from head and neck 5000, a prospective observational cohort of people with head and neck cancer Int. J. Cancer. 2018 143 1114 1127 10.1002/ijc.31416609936629607493 Search in Google Scholar

[6] Kobayashi K, Hisamatsu K, Suzui N, Hara A, Tomita H, Miyazaki T. A review of HPV-related head and neck cancer. J. Clin. Med. 2018; 7: 241. KobayashiK HisamatsuK SuzuiN HaraA TomitaH MiyazakiT A review of HPV-related head and neck cancer J. Clin. Med. 2018 7 241 10.3390/jcm7090241616286830150513 Search in Google Scholar

[7] Báez A. Genetic and environmental factors in head and neck cancer genesis. J Environ Sci Health C Environ Carcinog. Ecotoxicol. Rev. 2008; 26: 174–200. BáezA Genetic and environmental factors in head and neck cancer genesis J Environ Sci Health C Environ Carcinog. Ecotoxicol. Rev. 2008 26 174 200 10.1080/1059050080212943118569329 Search in Google Scholar

[8] Chuang SC, Jenab M, Heck JE, Bosetti C, Talamini R, Matsuo K, Castellsague X, Franceschi S, Herrero R, Winn DM, et al. Diet and the risk of head and neck cancer: A pooled analysis in the INHANCE consortium. Cancer Causes Control, 2012; 23: 69–88. ChuangSC JenabM HeckJE BosettiC TalaminiR MatsuoK CastellsagueX FranceschiS HerreroR WinnDM Diet and the risk of head and neck cancer: A pooled analysis in the INHANCE consortium Cancer Causes Control. 2012 23 69 88 10.1007/s10552-011-9857-x365440122037906 Search in Google Scholar

[9] Hashim D, Sartori S, Brennan P, Curado MP, Wünsch-Filho V, Divaris K, Olshan AF, Zevallos JP, Winn DM, Franceschi S, et al. The role of oral hygiene in head and neck cancer: Results from International Head and Neck Cancer Epidemiology (INHANCE) consortium. Ann. Oncol. 2016; 27: 1619–1625. HashimD SartoriS BrennanP CuradoMP Wünsch-FilhoV DivarisK OlshanAF ZevallosJP WinnDM FranceschiS The role of oral hygiene in head and neck cancer: Results from International Head and Neck Cancer Epidemiology (INHANCE) consortium Ann. Oncol. 2016 27 1619 1625 10.1093/annonc/mdw224495992927234641 Search in Google Scholar

[10] Marcu LG, Reid P, Bezak E. The promise of novel biomarkers for head and neck cancer from an imaging perspective. Int. J. Mol. Sci. 2018; 19: 2511. MarcuLG ReidP BezakE The promise of novel biomarkers for head and neck cancer from an imaging perspective Int. J. Mol. Sci. 2018 19 2511 10.3390/ijms19092511616511330149561 Search in Google Scholar

[11] Eze N, Lo YC, Burtness B. Biomarker driven treatment of head and neck squamous cell cancer. Cancers Head Neck. 2017; 2: 6. EzeN LoYC BurtnessB Biomarker driven treatment of head and neck squamous cell cancer Cancers Head Neck. 2017 2 6 10.1186/s41199-017-0025-1646053131093353 Search in Google Scholar

[12] Arantes LM, De Carvalho AC, Melendez ME, Carvalho AL. Serum, plasma and saliva biomarkers for head and neck cancer. Expert Rev. Mol. Diagn. 2018; 18: 85–112. ArantesLM De CarvalhoAC MelendezME CarvalhoAL Serum, plasma and saliva biomarkers for head and neck cancer Expert Rev. Mol. Diagn. 2018 18 85 112 10.1080/14737159.2017.140490629134827 Search in Google Scholar

[13] Dahiya K, Dhankhar R. Updated overview of current biomarkers in head and neck carcinoma. World J. Methodol. 2016; 6: 77–86. DahiyaK DhankharR Updated overview of current biomarkers in head and neck carcinoma World J. Methodol. 2016 6 77 86 10.5662/wjm.v6.i1.77480425427018324 Search in Google Scholar

[14] Arvelo F, Sojo F, Cotte C. Tumour progression and metastasis. Ecancermedicalscience, 2016; 10: 617. ArveloF SojoF CotteC Tumour progression and metastasis Ecancermedicalscience. 2016 10 617 10.3332/ecancer.2016.617475411926913068 Search in Google Scholar

[15] Nebbioso A, Tambaro FP, Dell’Aversana C, Altucci L. Cancer epigenetics: Moving forward. PLoS. Genet. 2018; 14: e1007362. NebbiosoA TambaroFP Dell’AversanaC AltucciL Cancer epigenetics: Moving forward PLoS. Genet. 2018 14 e1007362 10.1371/journal.pgen.1007362599166629879107 Search in Google Scholar

[16] Moore LD, Le T, Fan G. DNA methylation and its basic function. Neuropsychopharmacology. 2013; 38: 23–38. MooreLD LeT FanG DNA methylation and its basic function Neuropsychopharmacology. 2013 38 23 38 10.1038/npp.2012.112352196422781841 Search in Google Scholar

[17] Jin B, Robertson KD. DNA methyltransferases, DNA damage repair, and cancer. Adv. Exp. Med. Biol. 2013; 754: 3–29. JinB RobertsonKD DNA methyltransferases, DNA damage repair, and cancer Adv. Exp. Med. Biol. 2013 754 3 29 10.1007/978-1-4419-9967-2_1370727822956494 Search in Google Scholar

[18] Trimmer E: Methylenetetrahydrofolate reductase: Biochemical characterization and medical significance. Curr. Pharm. Des. 2013; 19: 2574–2593. TrimmerE Methylenetetrahydrofolate reductase: Biochemical characterization and medical significance Curr. Pharm. Des. 2013 19 2574 2593 10.2174/138161281131914000823116396 Search in Google Scholar

[19] Zhang H, Ma H, Li L, Zhang Z, Xu Y. Association of methylenetetra-hydrofolate dehydrogenase 1 polymorphisms with cancer: A meta-analysis. PLoS. One. 2013; 8: e69366. ZhangH MaH LiL ZhangZ XuY Association of methylenetetra-hydrofolate dehydrogenase 1 polymorphisms with cancer: A meta-analysis PLoS. One. 2013 8 e69366 10.1371/journal.pone.0069366371664323894459 Search in Google Scholar

[20] Banerjee RV, Matthews RG. Cobalamin-dependent methionine synthase. FASEB J. 1990; 4: 1450–1459. BanerjeeRV MatthewsRG Cobalamin-dependent methionine synthase FASEB J. 1990 4 1450 1459 10.1096/fasebj.4.5.24075892407589 Search in Google Scholar

[21] Yamada K, Gravel RA, Toraya T, Matthews RG. Human methionine synthase reductase is a molecular chaperone for human methionine synthase. Proc. Natl. Acad. Sci. USA. 2006; 103: 9476–9481. YamadaK GravelRA TorayaT MatthewsRG Human methionine synthase reductase is a molecular chaperone for human methionine synthase Proc. Natl. Acad. Sci. USA. 2006 103 9476 9481 10.1073/pnas.0603694103148043216769880 Search in Google Scholar

[22] Seetharam B, Li N. Transcobalamin II and its cell surface receptor. Vitam. Horm. 2000; 59: 337–366. SeetharamB LiN Transcobalamin II and its cell surface receptor Vitam. Horm. 2000 59 337 366 10.1016/S0083-6729(00)59012-8 Search in Google Scholar

[23] Pérez-Miguelsanz J, Vallecillo N, Garrido F, Reytor E, Pérez-Sala D, Pajares MA. Betaine homocysteine S-methyltransferase emerges as a new player of the nuclear methionine cycle. Biochim. Biophys. Acta. 2017; 1864: 1165–1182. Pérez-MiguelsanzJ VallecilloN GarridoF ReytorE Pérez-SalaD PajaresMA Betaine homocysteine S-methyltransferase emerges as a new player of the nuclear methionine cycle Biochim. Biophys. Acta. 2017 1864 1165 1182 10.1016/j.bbamcr.2017.03.00428288879 Search in Google Scholar

[24] Szegedi SS, Castro CC, Koutmos M, Garrow TA. Betaine-homocysteine S-methyltransferase-2 is an S-methylmethionine-homocysteine methyltransferase. J. Biol. Chem. 2008; 283: 8939–8945. SzegediSS CastroCC KoutmosM GarrowTA Betaine-homocysteine S-methyltransferase-2 is an S-methylmethionine-homocysteine methyltransferase J. Biol. Chem. 2008 283 8939 8945 10.1074/jbc.M710449200227637418230605 Search in Google Scholar

[25] Jhee KH, Kruger WD. The role of cystathionine β-synthase in homocysteine metabolism. Antioxid. Redox. Signal. 2005; 7: 813–822. JheeKH KrugerWD The role of cystathionine β-synthase in homocysteine metabolism Antioxid. Redox. Signal. 2005 7 813 822 10.1089/ars.2005.7.81315890029 Search in Google Scholar

[26] Quadros EV, Sequeira JM. Cellular uptake of cobalamin: Transcobalamin and the TCblR/CD320 receptor. Biochimie. 2013; 95: 1008–1018. QuadrosEV SequeiraJM Cellular uptake of cobalamin: Transcobalamin and the TCblR/CD320 receptor Biochimie. 2013 95 1008 1018 10.1016/j.biochi.2013.02.004390248023415653 Search in Google Scholar

[27] Allen LH. Vitamin B-12. Adv.. Nutr. 2012; 3: 54–55. AllenLH Vitamin B-12 Adv.. Nutr. 2012 3 54 55 10.3945/an.111.001370326261422332101 Search in Google Scholar

[28] Brunaud L, Alberto JM, Ayav A, Gérard P, Namour F, Antunes L, Braun M, Bronowicki JP, Bresler L, Guéant JL. Vitamin B12 is a strong determinant of low methionine synthase activity and DNA hypomethylation in gastrectomized rats. Digestion. 2003; 68: 133–140. BrunaudL AlbertoJM AyavA GérardP NamourF AntunesL BraunM BronowickiJP BreslerL GuéantJL Vitamin B12 is a strong determinant of low methionine synthase activity and DNA hypomethylation in gastrectomized rats Digestion. 2003 68 133 140 10.1159/00007530714646334 Search in Google Scholar

[29] Castro R, Barroso M, Rocha M, Esse R, Ramos R, Ravasco P, Rivera I, de Almeida IT. The TCN2 776C>G polymorphism correlates with vitamin B₁₂ cellular delivery in healthy adult populations. Clin. Biochem. 2010; 43: 645–649. CastroR BarrosoM RochaM EsseR RamosR RavascoP RiveraI de AlmeidaIT The TCN2 776C>G polymorphism correlates with vitamin B₁₂ cellular delivery in healthy adult populations Clin. Biochem. 2010 43 645 649 10.1016/j.clinbiochem.2010.01.01520144600 Search in Google Scholar

[30] Stanisławska-Sachadyn A, Woodside JV, Sayers CM, Yarnell JW, Young IS, Evans AE, Mitchell LE, Whitehead AS. The transcobalamin (TCN2) 776C>G polymorphism affects homocysteine concentrations among subjects with low vitamin B₁₂ status. Eur. J. Clin. Nutr. 2010; 64: 1338–1343. Stanisławska-SachadynA WoodsideJV SayersCM YarnellJW YoungIS EvansAE MitchellLE WhiteheadAS The transcobalamin (TCN2) 776C>G polymorphism affects homocysteine concentrations among subjects with low vitamin B₁₂ status Eur. J. Clin. Nutr. 2010 64 1338 1343 10.1038/ejcn.2010.15720808328 Search in Google Scholar

[31] Malinowska K, Morawiec-Sztandera A, Majsterek I, Kaczmarczyk D. TC2 C776G polymorphism studies in patients with oral cancer in the Polish population. Pol. J. Pathol. 2016; 67: 277–282. MalinowskaK Morawiec-SztanderaA MajsterekI KaczmarczykD TC2 C776G polymorphism studies in patients with oral cancer in the Polish population Pol. J. Pathol. 2016 67 277 282 10.5114/pjp.2016.6378028155977 Search in Google Scholar

[32] Cheng J, Zhu WL, Dao JJ, Li SQ, Li Y. Relationship between polymorphism of methylenetetrahydrofolate dehydrogenase and congenital heart defect. Biomed. Environ. Sci. 2005; 18: 58–64. ChengJ ZhuWL DaoJJ LiSQ LiY Relationship between polymorphism of methylenetetrahydrofolate dehydrogenase and congenital heart defect Biomed. Environ. Sci. 2005 18 58 64 Search in Google Scholar

[33] Pietrzyk JJ, Bik-Multanowski M. 776C>G polymorphism of the trans-cobalamin II gene as a risk factor for spinabifida. Mol. Genet. Metab. 2003; 80: 364. PietrzykJJ Bik-MultanowskiM 776C>G polymorphism of the trans-cobalamin II gene as a risk factor for spinabifida Mol. Genet. Metab. 2003 80 364 10.1016/S1096-7192(03)00131-8 Search in Google Scholar

[34] Lyman GH. Counting the costs of cancer care. Lancet Oncol. 2013; 14: 1142–1143. LymanGH Counting the costs of cancer care Lancet Oncol. 2013 14 1142 1143 10.1016/S1470-2045(13)70480-7 Search in Google Scholar

[35] Alho OP, Teppo H, Mäntyselkä P, Kantola S. Head and neck cancer in primary care: Presenting symptoms and the effect of delayed diagnosis of cancer cases. CMAJ, 2006; 174: 779–784. AlhoOP TeppoH MäntyselkäP KantolaS Head and neck cancer in primary care: Presenting symptoms and the effect of delayed diagnosis of cancer cases CMAJ. 2006 174 779 784 10.1503/cmaj.050623140239416534084 Search in Google Scholar

[36] Makni L, Zidi S, Barbiroud M, Ahmed AB, Gazouani E, Mezlini A, Stayoussef M, Yacoubi-Loueslati B. Increased risks between TLR2 (-196 to -174 ins/del) and TLR3 1377C>T variants and head and neck cancers in Tunisia. Centr. Eur. J. Immunol. 2019; 44: 144–149. MakniL ZidiS BarbiroudM AhmedAB GazouaniE MezliniA StayoussefM Yacoubi-LoueslatiB Increased risks between TLR2 (-196 to -174 ins/del) and TLR3 1377C>T variants and head and neck cancers in Tunisia Centr. Eur. J. Immunol. 2019 44 144 149 10.5114/ceji.2019.87065674554931530984 Search in Google Scholar

[37] Demokan S, Dalay N. Role of DNA methylation in head and neck cancer. Clin. Epigenetics. 2011; 2: 123–150. DemokanS DalayN Role of DNA methylation in head and neck cancer Clin. Epigenetics. 2011 2 123 150 10.1007/s13148-011-0045-3336539122704334 Search in Google Scholar

[38] Jeziorska DM, Murray RJ, De Gobbi M, Gaentzsch R, Garrick D, Ayyub H, Chen T, Li E, Telenius J, Lynch M, et al. DNA methylation of intragenic CpG islands depends on their transcriptional activity during differentiation and disease. Proc. Natl. Acad. Sci. USA. 2017; 114: E7526–E7535. JeziorskaDM MurrayRJ De GobbiM GaentzschR GarrickD AyyubH ChenT LiE TeleniusJ LynchM DNA methylation of intragenic CpG islands depends on their transcriptional activity during differentiation and disease Proc. Natl. Acad. Sci. USA. 2017 114 E7526 E7535 10.1073/pnas.1703087114559464928827334 Search in Google Scholar

[39] Hazra A, Fuchs CS, Kawasaki T, Kirkner GJ, Hunter DJ, Ogino S. Germline polymorphisms in the one-carbon metabolism pathway and DNA methylation in colorectal cancer. Cancer Causes Control. 2010; 21: 331–345. HazraA FuchsCS KawasakiT KirknerGJ HunterDJ OginoS Germline polymorphisms in the one-carbon metabolism pathway and DNA methylation in colorectal cancer Cancer Causes Control. 2010 21 331 345 10.1007/s10552-009-9464-2357097819936946 Search in Google Scholar

[40] Krakowczyk Ł, Strzelczyk JK. Epigenetic modification of gene expression in colorectal carcinogenesis. Współcz Onkol. 2007; 11: 289–294. KrakowczykŁ StrzelczykJK Epigenetic modification of gene expression in colorectal carcinogenesis Współcz Onkol. 2007 11 289 294 Search in Google Scholar

[41] Zara-Lopes T, Galbiatti-Dias AL, Castanhole-Nunes MM, Padovani-Júnior JA, Maniglia JV, Pavarino EC, Goloni-Bertollo EM. Polymorphisms in MTHFR, MTR, RFC1 and CβS genes involved in folate metabolism and thyroid cancer: A case-control study. Arch. Med. Sci. 2019; 15: 522–530. Zara-LopesT Galbiatti-DiasAL Castanhole-NunesMM Padovani-JúniorJA ManigliaJV PavarinoEC Goloni-BertolloEM Polymorphisms in MTHFR, MTR, RFC1 and CβS genes involved in folate metabolism and thyroid cancer: A case-control study Arch. Med. Sci. 2019 15 522 530 10.5114/aoms.2018.73091642520730899306 Search in Google Scholar

[42] De Castro TB, Rodrigues-Fleming GH, Oliveira-Cucolo JG, Silva JN, Silva FP, Raposo LS, Maniglia JV, Pavarino EC, Batista Arantes LM, Galbiatti-Dias AL, et al. Gene polymorphisms involved in folate metabolism and DNA methylation with the risk of head and neck cancer. Asian Pac. J. Cancer Prev. 2020; 21: 3751–3759. De CastroTB Rodrigues-FlemingGH Oliveira-CucoloJG SilvaJN SilvaFP RaposoLS ManigliaJV PavarinoEC Batista ArantesLM Galbiatti-DiasAL Gene polymorphisms involved in folate metabolism and DNA methylation with the risk of head and neck cancer Asian Pac. J. Cancer Prev. 2020 21 3751 3759 10.31557/APJCP.2020.21.12.3751804629433369477 Search in Google Scholar

[43] Cai CQ, Fang YL, Shu JB, Zhao LS, Zhang RP, Cao LR, Wang YZ, Zhi XF, Cui HL, Shi OY, Liu W. Association of neural tube defects with maternal alterations and genetic polymorphisms in one-carbon metabolic pathway. Ital. J. Pediatr. 2019; 45: 37. CaiCQ FangYL ShuJB ZhaoLS ZhangRP CaoLR WangYZ ZhiXF CuiHL ShiOY LiuW Association of neural tube defects with maternal alterations and genetic polymorphisms in one-carbon metabolic pathway Ital. J. Pediatr. 2019 45 37 10.1186/s13052-019-0630-1641684230867013 Search in Google Scholar

[44] Jiang J, Zhang Y, Wei L, Sun Z, Liu Z. Association between MTHFD1 G1958A polymorphism and neural tube defects susceptibility: A meta-analysis. PLoS One. 2014; 9: e101169. JiangJ ZhangY WeiL SunZ LiuZ Association between MTHFD1 G1958A polymorphism and neural tube defects susceptibility: A meta-analysis PLoS One. 2014 9 e101169 10.1371/journal.pone.0101169407626424977710 Search in Google Scholar

[45] Parle-McDermott A, Pangilinan F, Mills JL, Signore CC, Molloy AM, Cotter A, Conley M, Cox C, Kirke PN, Scott JM, Brody LC. A polymorphism in the MTHFD1 gene increases a mother’s risk of having an unexplained second trimester pregnancy loss. Mol. Hum. Reprod. 2005; 11: 477–480. Parle-McDermottA PangilinanF MillsJL SignoreCC MolloyAM CotterA ConleyM CoxC KirkePN ScottJM BrodyLC A polymorphism in the MTHFD1 gene increases a mother’s risk of having an unexplained second trimester pregnancy loss Mol. Hum. Reprod. 2005 11 477 480 10.1093/molehr/gah20416123074 Search in Google Scholar

[46] Xuan C, Li H, Zhao JX, Wang HW, Wang Y, Ning CP, Liu Z, Zhang BB, He GW, Lun LM. Association between MTHFR polymorphisms and congenital heart disease: A meta-analysis based on 9,329 cases and 15,076 controls. Sci. Rep. 2014; 4: 7311. XuanC LiH ZhaoJX WangHW WangY NingCP LiuZ ZhangBB HeGW LunLM Association between MTHFR polymorphisms and congenital heart disease: A meta-analysis based on 9,329 cases and 15,076 controls Sci. Rep. 2014 4 7311 10.1038/srep07311425518825472587 Search in Google Scholar

[47] Kempisty B, Sikora J, Lianeri M, Szczepankiewicz A, Czerski P, Hauser J, Jagodzinski PP. MTHFD 1958G>A and MTR 2756A>G polymorphisms are associated with bipolar disorder and schizophrenia. Psychiatr. Genet. 2007; 17: 177–181. KempistyB SikoraJ LianeriM SzczepankiewiczA CzerskiP HauserJ JagodzinskiPP MTHFD 1958G>A and MTR 2756A>G polymorphisms are associated with bipolar disorder and schizophrenia Psychiatr. Genet. 2007 17 177 181 10.1097/YPG.0b013e328029826f17417062 Search in Google Scholar

[48] Sutherland HG, Hermile H, Sanche R, Menon S, Lea RA, Haupt LM, Griffiths LR. Association study of MTHFD1 coding polymorphisms R134K and R653Q with migraine susceptibility. Headache. 2014; 54: 1506–1514. SutherlandHG HermileH SancheR MenonS LeaRA HauptLM GriffithsLR Association study of MTHFD1 coding polymorphisms R134K and R653Q with migraine susceptibility Headache. 2014 54 1506 1514 10.1111/head.1242825039261 Search in Google Scholar

[49] Stevens VL, McCullough ML, Pavluck AL, Talbot JT, Feigelson HS, Thun MJ, Calle EE. Association of polymorphisms in one-carbon metabolism genes and postmenopausal breast cancer incidence. Cancer Epidemiol. Biomarkers Prev. 2007; 16: 1140–1147. StevensVL McCulloughML PavluckAL TalbotJT FeigelsonHS ThunMJ CalleEE Association of polymorphisms in one-carbon metabolism genes and postmenopausal breast cancer incidence Cancer Epidemiol. Biomarkers Prev. 2007 16 1140 1147 10.1158/1055-9965.EPI-06-103717548676 Search in Google Scholar

[50] Li SY, Rong M, Iacopetta B. Germ-line variants in methyl-group metabolism genes and susceptibility to DNA methylation in human breast cancer. Oncol. Rep. 2006; 15: 221–225. LiSY RongM IacopettaB Germ-line variants in methyl-group metabolism genes and susceptibility to DNA methylation in human breast cancer Oncol. Rep. 2006 15 221 225 10.3892/or.15.1.221 Search in Google Scholar

[51] Wang L, Ke Q, Chen W, Wang J, Tan Y, Zhou Y, Hua Z, Ding W, Niu J, Shen J, et al. Polymorphisms of MTHFD, plasma homocysteine levels, and risk of gastric cancer in a high-risk Chinese population. Clin. Cancer Res. 2007; 13: 2526–2532. WangL KeQ ChenW WangJ TanY ZhouY HuaZ DingW NiuJ ShenJ Polymorphisms of MTHFD, plasma homocysteine levels, and risk of gastric cancer in a high-risk Chinese population Clin. Cancer Res. 2007 13 2526 2532 10.1158/1078-0432.CCR-06-229317438114 Search in Google Scholar

[52] Krajinovic M, Lemieux-Blanchard É, Chiasson S, Primeau M, Costea I, Moghrabi A. Role of polymorphism in MTHFR and MTHFD1 genes in the outcome of childhood acute lymphoblastic leukemia. Pharmacogenomics J. 2004; 4: 66–72. KrajinovicM Lemieux-BlanchardÉ ChiassonS PrimeauM CosteaI MoghrabiA Role of polymorphism in MTHFR and MTHFD1 genes in the outcome of childhood acute lymphoblastic leukemia Pharmacogenomics J. 2004 4 66 72 10.1038/sj.tpj.650022414647408 Search in Google Scholar

[53] Liu G, Qi C, Xu Q, Wu B, Wang Y, Xue C. Lack of association between methylenetetrahydrofolate dehydrogenase 1 G1958A polymorphism and prostate cancer risk: A meta-analysis. Tumor Biol. 2014; 35: 2029–2033. LiuG QiC XuQ WuB WangY XueC Lack of association between methylenetetrahydrofolate dehydrogenase 1 G1958A polymorphism and prostate cancer risk: A meta-analysis Tumor Biol. 2014 35 2029 2033 10.1007/s13277-013-1269-y24197977 Search in Google Scholar

[54] da Silva LM, Silva JN, Galbiatti AL, Succi M, Ruiz MT, Raposo LS, Maniglia JV, Pavarino-Bertelli EC, Goloni-Bertollo EM. Head and neck carcinogenesis: Impact of MTHFD1 G1958A polymorphism. Rev. Assoc. Med. Bras. 2011; 57: 194–199. da SilvaLM SilvaJN GalbiattiAL SucciM RuizMT RaposoLS ManigliaJV Pavarino-BertelliEC Goloni-BertolloEM Head and neck carcinogenesis: Impact of MTHFD1 G1958A polymorphism Rev. Assoc. Med. Bras. 2011 57 194 199 Search in Google Scholar

[55] Hazra A, Wu K, Kraft P, Fuchs CS, Giovannucci EL, Hunter DJ. Twenty-four non-synonymous polymorphisms in the one-carbon metabolic pathway and risk of colorectal adenoma in the Nurses’ Health Study. Carcinogenesis. 2007; 28: 1510–1519. HazraA WuK KraftP FuchsCS GiovannucciEL HunterDJ Twenty-four non-synonymous polymorphisms in the one-carbon metabolic pathway and risk of colorectal adenoma in the Nurses’ Health Study Carcinogenesis. 2007 28 1510 1519 10.1093/carcin/bgm06217389618 Search in Google Scholar

[56] Zheng S, Wu C, Yang W, Xia X, Lin X, Jiang L, Ding R, Jiang Y. An analysis of transcobalamin II gene polymorphisms and serum levels of homocysteine, folate and vitamin B₁₂ in Chinese patients with Crohn’s disease. Dig. Dis. 2017; 35: 463–471. ZhengS WuC YangW XiaX LinX JiangL DingR JiangY An analysis of transcobalamin II gene polymorphisms and serum levels of homocysteine, folate and vitamin B₁₂ in Chinese patients with Crohn’s disease Dig. Dis. 2017 35 463 471 10.1159/00047184828472811 Search in Google Scholar

[57] Green R. Peripheral neuropathy risk and a transcobalamin polymorphism: Connecting the dots between excessive folate intake and disease susceptibility. Am. J. Clin. Nutr. 2016; 104: 1495–1496. GreenR Peripheral neuropathy risk and a transcobalamin polymorphism: Connecting the dots between excessive folate intake and disease susceptibility Am. J. Clin. Nutr. 2016 104 1495 1496 10.3945/ajcn.116.146365511873727852614 Search in Google Scholar

[58] Oussalah A, Levy J, Filhine-Trésarrieu P, Namour F, Guéant JL. Association of TCN2 rs1801198 c.776G>C polymorphism with markers of one-carbon metabolism and related diseases: A systematic review and meta-analysis of genetic association studies. Am. J. Clin. Nutr. 2017; 106: 1142–1156. OussalahA LevyJ Filhine-TrésarrieuP NamourF GuéantJL Association of TCN2 rs1801198 c.776G>C polymorphism with markers of one-carbon metabolism and related diseases: A systematic review and meta-analysis of genetic association studies Am. J. Clin. Nutr. 2017 106 1142 1156 10.3945/ajcn.117.156349561178328814397 Search in Google Scholar

Postępy Higieny i Medycyny Doświadczalnej

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

Data publikacji: 06 Apr 2022

Tom & Zeszyt: Tom 76 (2022) - Zeszyt 1 (January 2022)

Zakres stron: 30 - 38

Otrzymano: 07 Feb 2021

Przyjęty: 07 Sep 2021

Postępy Higieny i Medycyny Doświadczalnej

Keywords

Fig. 1

Fig. 2

Fig. 1

Fig. 2

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

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

Detailed characterization of MTHFD1 and TCN2 gene polymorphisms

The primers, length of PCR products, and restriction enzymes

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

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