Genetics of Non-Communicable Disease Research Center, Zahedan University of Medical Sciences, Zahedan, Iran

Foundation: The current review was expected to look at the conceivable relationship between methylene tetrahydrofolate reductase (MTHFR) quality polymorphisms and youth intense lymphoblastic leukemia (ALL) in an example of Iranian populace. Subjects and Techniques: A sum of 220 subjects incorporating 100 youngsters determined to have ALL and 120 solid kids partook for the situation control study. The single nucleotide polymorphisms (SNPs) of not entirely settled by ARMS-PCR or PCR-RFLP strategy.

Conclusion: The ongoing examination discoveries showed that MTHFR rs1801131 and rs13306561 polymorphisms diminished the gamble of ALL in the populace which has been examined. Further examinations with bigger test sizes and various nationalities are expected to approve our discoveries.

Keywords: MTHFR, Polymorphism, Intense lymphocytic leukemia

Children who reside in wealthy nations are currently most likely to die from paediatric cancer. The most frequent childhood malignancy, acute lymphoblastic leukaemia (ALL), makes up about 26.8% of all paediatric malignancies in children under the age of 15. Although the cause of ALL is mostly unknown, genetic diversity is known to play a significant influence in the progression of the disease.3,4 The peak rate of childhood ALL occurs between the ages of 2 and 5, suggesting that ALL may begin in utero or during the neonatal stage.

The short arm of chromosome 1 is where the MTHFR gene is found (1p36.3). A vital enzyme in the metabolism of folate, 5 MTHFR protein catalyses the irreversible conversion of 5, 10-5-Methyltetrahydrofolate (5, 10-MTHF) is converted to 5-Methyl THF. The primary circulating form of folate is 5-methyl THF, which supplies the methyl group necessary for homocysteine methylation and conversion to methionine. Additionally, it contributes to the synthesis of Sadenosylmethionine (SAM), the all-purpose donor of one carbon atom for the methylation of many substrates. 6-8 There are two well-known polymorphisms in the MTHFR gene: First, there is a C to T transition at nucleotide 677 (C677T, rs1801133), which results in the substitution of alanine with valine at codon 222, lowers enzyme activity, and ultimately raises homocysteine levels and changes the distribution of folate. 9 The second is the A to C transition at position 1298 (A1298C, rs1801131), which changes glutamate to alanine at codon position 429 and likewise causes MTHFR to be less active.

The connections between ALL and MTHFR gene polymorphisms have been examined in a number of genetic association studies, however the findings have been conflicting or ambiguous in different groups. 11-14 The current investigation sought to determine whether there may be a link between ALL risk and the MTHFR gene polymorphisms rs1801131 (A1298C), rs1801133 (C677T), and rs13306561 in a sample of the Iranian population.

Patients This case-control concentrate on has been finished in youngsters determined to have ALL (n=100) and control people (n=120), who were matched for age and sex in southeast of Iran (Zahedan). The review plan and the enrolment methodology have been depicted previously.15-18 Neighborhood morals advisory group of Zahedan College of Clinical Sciences was supported the project and informed assent was acquired from guardians of cases and controls (Moral code: 6858).

DNA was extricated from fringe entire blood utilizing salting-out method.19 Genotyping Genotyping of MTHFR rs1801133 (C677T,Ala222Val) and rs13306561 quality polymorphisms was finished by PCR-RFLP technique. For rs1801133 variation, the forward and switch preliminaries were 5′-ACTCTCTCTGCCCAGTCCCTGTG-3′ and 5′ AAGAACTCAGCGAACTCAGCACT-3′, individually.In each 0.20 ml PCR response tube, 1 µl of genomic DNA (~100 ng/ml), 1 µl of every preliminary, 10 µl of 2x Prime Taq Premix (Genet Bio, Korea) and 7 µl ddH2O were added. The PCR conditions were set as follows: 95˚C for 5 min, 30 patterns of 95˚C for 30s, 64˚C for 30s and 72 ˚C for 30s and a last expansion step of 72 ˚C for 10 min. Then, 10 µl of PCR item is processed with HinfI limitation chemical.

The Tallele was processed and created 222-bp and 167-bp sections, while the C allele was undigested and delivered 389-bp item. For MTHFR rs13306561 polymorphism, a bunch of forward and switch preliminary matches was 5′-GGCCGAGCGTTCTGAGTC-3′ and 5′-CACTAATCCCGCGAAGGGTGCTCA-3′, individually. The PCR cycling conditions were the underlying denaturation step at 95 °C for 5 min, trailed by 30 patterns of 95 °C for 30s, 64 °C for 30s, and 72 °C for 30s with a last expansion of 72 °C for 10 min. Ten microliters of the PCR item were processed by HpyF3I limitation catalyst. The C allele was processed and delivered 162-bp and 23-bp, while the T allele was undigested (185-bp).Genotyping of MTHFR rs1801131 (A1298C) polymorphism was finished utilizing allele explicit polymerase chain response (AS-PCR).

The groundwork arrangements were: conventional roundworks, 5′- AGACCTTCCTTGCAAATACAT-3′; An allele, 5′-GAAGACTTCAAAGACACTCT-3′; C allele, and 5′-GAAGACTTCAAAGACACTCG-3′.The beta-2 microglobulin (B2MF: 5′- TGTAAACACTTGGTGCCTGATATAGCTTGA-3′, B2MR:5′-CATCAGTATCTCAGCAGGTGCCACTAATCT-3′) was utilized as inside control.The PCR cycling conditions were 95˚C for 5 min, 33 patterns of 95˚C for 30s, 55˚C for 20s, and 72˚C for 30s, trailed by a last expansion step for 10 min at 72˚C. The items size were 317-bp for both of the An or C alleles and 574-bp for the inner control.

Software from the statistical package SPSS 20 was used for the statistical analysis. Independent sample t-test and 2 test were used to assess the data. Polymorphisms in the MTHFR gene are linked and the odds ratio (OR) and 95% confidence intervals (95%CI) from logistic regression analysis were calculated. SNPStats software was used to conduct a haplotype analysis. 20 A p-value of 0.05 or less was regarded as statistically significant.

Everything is a multifactorial infection impacted by hereditary and ecological elements. In the ongoing review, we analyzed the effect of MTHFR quality polymorphisms on the gamble of experience growing up ALL in an example of the Iranian populace. Our discoveries demonstrated that rs1801131 (A1298C) as well as rs13306561 T>C variations in MTHFR fundamentally diminished the gamble of ALL in our populace. While, no critical relationship between the MTHFR C677T (rs1801133) variation and Everything was found. In concurrence with our finding various past examinations have tracked down no critical relationship between MTHFR C677T variation and hazard of experience growing up ALL.21-24 Though, not many gatherings have detailed that MTHFR C677T variation decreased25 or expanded the gamble of ALL.12 Our outcomes showed similar discoveries in regards to the job of MTHFR A1298C in the reduction of the ALL risk.12 rather than our discoveries, no critical relationship between MTHFR A1298C polymorphism and ALL has been found.26-29 A few examinations recommended that MTHFR A1298C polymorphism expanded the gamble of ALL.22,30-31 As of late, a meta-investigation research has been finished in a Chinese populace and its outcomes proposed that both MTHFR C677T and A1298C polymorphisms might be possible biomarkers for ALL risk[13].

Generally speaking, a huge diminished affiliation was found between the MTHFR C677T polymorphism and ALL gamble. Conversely, C677T and A1298C variations were essentially connected with an expanded gamble of ALL in clinic based studies,which included people with obscure identity. Dong et al.14 have done a meta-investigation and showed that MTHFR rs1801131 CC genotype polymorphism essentially expanded the gamble of myeloid leukemia in Asian populaces. Likewise, Jiang et al.25 have additionally demonstrated that 677TT genotype diminished the gamble of ALL in Caucasians in a meta-examination study (OR=0.715, 95%CI=0.655-0.781, p=0.000).

Be that as it may, no critical affiliation was found between C677T polymorphism and ALL in Asians and others. In another meta-examination study, Yan et al.32 have found a general relationship between 677T variation genotypes and decreased youth ALL gamble, however they found no critical relationship between MTHFR A1298C polymorphism and adolescence ALL gamble. In another meta-examination, it has been showed that the 677T allele was defensive, while A1298C was related with peripheral expansion in the gamble of life as a youngster ALL.30 It has been accounted for that people conveying MTHFR 677T allele might have a higher relative gamble of pediatric ALL mortality.33 In a meta-investigation study, no affiliation has been accounted for between MTHFR A1298C and C677T polymorphisms and pediatric ALL.29 As of late, it has likewise been shown that the gamble of ALL improvement in kids with MTHFR 677 TT genotype diminished in Asian population.34 Atashrazm et al.35 researched the effect of MTHFR C677T and A1298C polymorphisms and the gamble of ALL in patients alluded to the Iranian Blood Bonding Association (Tehran, Iran). In concurrence with our finding, they tracked down no huge relationship between MTHFR C677T variation and the gamble of life as a youngster ALL. Strangely, they likewise tracked down no relationship between MTHFR A1298C polymorphism and the gamble of ALL. It is essential to make reference to that the hereditary foundation of our populace (Baluch and Sistani) was differentfrom their populace, which utilized in their review

(patients alluded to the Iranian Blood Bonding Association, Tehran, Iran who had different hereditary foundation). Accordingly, it very well may be presumed that the information with respect to the effect of MTHFR on youth ALL are conflicting. There is no great explanation for the various outcomes in assorted examinations. Ethnic, hereditary, and additionally ecological variables as well as quality eating regimen communication might connect in different ways to one or the other reduction or increment the gamble of life as a youngster ALL in various regions. Taken together, our discoveries showed that MTHFR rs1801131 and rs13306561 polymorphisms diminished the gamble of ALL in an example of Iranian populace. To additionally affirm these discoveries, bigger example sizes with different nationalities are required.

The results of the current case-control study showed an association between MTHFR rs1801131 and rs13306561 polymorphisms and risk of ALL in a sample of Iranian population.

1. Siegel R, Naishadham D, Jemal A. Cancer statistics,2013. CA Cancer J Clin 2013; 63(1):11-30.

2. Kaatsch P. Epidemiology of childhood cancer. Cancer Treat Rev. 2010; 36(4):277-85.

3. Guo LM, Xi JS, Ma Y, et al. ARID5B gene rs10821936 polymorphism is associated with childhood acute lymphoblastic leukemia: a meta-analysis based on 39,116 subjects. Tumour Biol. 2014; 35(1):709-13.

4. Ma Y, Sui Y, Wang L, et al. Effect of GSTM1 null genotype on risk of childhood acute leukemia: a metaanalysis. Tumour Biol. 2014; 35(1):397-402.

5. Wiemels JL, Smith RN, Taylor GM, et al.Methylenetetrahydrofolate reductase (MTHFR) polymorphisms and risk of molecularly defined subtypes of childhood acute leukemia. Proc Natl Acad Sci U S A.2001; 98(7):4004-9.

6. Franco RF, Simoes BP, Tone LG, et al. The methylenetetrahydrofolate reductase C677T gene polymorphism decreases the risk of childhood acute lymphocytic leukaemia. Br J Haematol. 2001; 115(3):616-8.

7. De Jonge R, Tissing WJ, Hooijberg JH, et al.Polymorphisms in folate-related genes and risk of pediatric acute lymphoblastic leukemia. Blood. 2009; 113(10):2284-9.

8. Sohn KJ, Jang H, Campan M, et al. The methylenetetrahydrofolate reductase C677T mutation induces cell-specific changes in genomic DNA methylation and uracil misincorporation: a possible molecular basis for the site-specific cancer risk modification. Int J Cancer. 2009; 124(9):1999-2005.

9. Frosst P, Blom HJ, Milos R, et al. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet. 1995; 10(1):111-3.

10. Van der Put NM, Gabreels F, Stevens EM, et al. A second common mutation in the methylenetetrahydrofolate reductase gene: an additional risk factor for neural-tube defects? Am J Hum Genet. 1998; 62(5):1044-51.

11. Kreile M, Rots D, Piekuse L, et al. Lack of Association between Polymorphisms in Genes MTHFR and MDR1 with Risk of Childhood Acute Lymphoblastic Leukemia.Asian Pac J Cancer Prev. 2014; 15(22):9707-11.

12. Silva RM, Fontes AC, Silva KA, et al. Polymorphisms involved in folate metabolism pathways and the risk of the development of childhood acute leukemia. Genet Test Mol Biomarkers. 2013; 17(2):147-52.

13. Xiao Y, Deng TR, Su CL, et al.Methylenetetrahydrofolate reductase polymorphisms and susceptibility to acute lymphoblastic leukemia in a chinese population: a meta-analysis. Oncol Res Treat. 2014; 37(10):576-82.

14. Dong S, Liu Y, Chen J. MTHFR gene polymorphism and risk of myeloid leukemia: a meta-analysis. Tumour Biol. 2014; 35(9):8913-9.

15. Hashemi M, Sheybani-Nasab M, Naderi M, et al. Association of functional polymorphism at the miR-502- binding site in the 3' untranslated region of the SETD8 gene with risk of childhood acute lymphoblastic leukemia, a preliminary report. Tumour Biol. 2014; 35(10):10375-9.

16. Hasani SS, Hashemi M, Eskandari-Nasab E, et al. A functional polymorphism in the miR-146a gene is associated with the risk of childhood acute lymphoblastic leukemia: a preliminary report. Tumour Biol. 2014;35(1):219-25.

17. Hashemi M, Ebrahimi M, Amininia S, et al. Evaluation of rs3102735 and rs2073617 Osteoprotegerin Gene Polymorphisms and the Risk of Childhood Acute lymphoblastic Leukemia in Zahedan Southeast Iran. Int J Hematol Oncol Stem Cell Res. 2014; 8(4):39-44.

18. Bahari G, Hashemi M, Naderi M, et al. IKZF1 gene polymorphisms increased the risk of childhood acute lymphoblastic leukemia in an Iranian population. Tumour Biol. 2016. DOI: 10.1007/s13277-016-4853-0.

19. Hashemi M, Hanafi Bojd H, Eskandari Nasab E, et al. Association of Adiponectin rs1501299 and rs266729 Gene Polymorphisms With Nonalcoholic Fatty Liver Disease. Hepat Mon. 2013; 13(5):e9527.

20. Sole X, Guino E, Valls J, et al. SNPStats: a web tool for the analysis of association studies. Bioinformatics. 2006; 22(15):1928-9.

21. Balta G, Yuksek N, Ozyurek E, et al. Characterization of MTHFR, GSTM1, GSTT1, GSTP1, and CYP1A1 genotypes in childhood acute leukemia. Am J Hematol. 2003; 73(3):154-60.

22. Li X, Liao Q, Zhang S, et al. Association of methylenetetrahytrofolate reductase (MTHFR) C677T and A1298C polymorphisms with the susceptibility of childhood acute lymphoblastic leukaemia (ALL) in Chinese population. Eur J Med Res. 2014; 19:5.

23. Azhar MR, Rahimi Z, Vaisi-Raygani A, et al. Lack of association between MTHFR C677T and A1298C polymorphisms and risk of childhood acute lymphoblastic leukemia in the Kurdish population from Western Iran.Genet Test Mol Biomarkers. 2012; 16(3):198-202.

24. Jiang N, Zhu X, Zhang H, et al. The relationship between ethylenetetrahydrofolate reductase polymorphism and hematological malignancy. Clin Lab. 2014; 60(5):767-74.

25. Jiang Y, Hou J, Zhang Q, et al. The MTHFR C677T polymorphism and risk of acute lymphoblastic leukemia:an updated meta-analysis based on 37 case-control studies. Asian Pac J Cancer Prev. 2013; 14(11):6357-62.

26. Bellampalli R, Phani NM, Bhat KG, et al. Significance of 5,10 ethylenetetrahydrofolate reductase gene variants in acute lymphoblastic leukemia in Indian population: an experimental, computational and metaanalysis. Leuk Lymphoma. 2015; 56(5):1450-9.

27. Sazawal S, Chaubey R, Kaur P, et al. MTHFR Gene Polymorphisms and the Risk of Acute Lymphoblastic Leukemia in Adults and Children: A Case Control Study in India. Indian J Hematol Blood Transfus. 2014; 30(4):219-25.

28. Schnakenberg E, Mehles A, Cario G, et al. Polymorphisms of ethylenetetrahydrofolate reductase (MTHFR) and susceptibility to pediatric acute lymphoblastic leukemia in a German study population. BMC Med Genet. 2005; 6:23.

29. Zintzaras E, Doxani C, Rodopoulou P, et al. Variants of the MTHFR gene and susceptibility to acute lymphoblastic leukemia in children: a synthesis of genetic association studies. Cancer Epidemiol. 2012; 36(2):169-76.

30. Roy Moulik N, Parveen F, Kumar A, et al. MTHFR gene polymorphism in acute lymphoblastic leukemia among North Indian children: a case-control study and metaanalysis updated from 2011. J Hum Genet. 2014;59(7):397-404.

31. Kim NK, Chong SY, Jang MJ, et al. Association of the methylenetetrahydrofolate reductase polymorphism in Korean patients with childhood acute lymphoblastic leukemia. Anticancer Res. 2006; 26(4B):2879-81.

32. Yan J, Yin M, Dreyer ZE, et al. A meta-analysis of MTHFR C677T and A1298C olymorphisms and risk of IJHOSCR, 1 July 2016. Volume 10

Taheri Mohsen . Methylenetetrahydrofolate Reductase (MTHFR) Gene Polymorphisms with Childhood Acute Lymphoblastic Leukemia Susceptibility in an Iranian Population. World Journal Of Hematology And Oncology 2022.