Association Between Tumor Necrosis Factor Alpha Gene Polymorphism (-376) and Multiple Sclerosis in Turkish Population

Öz

The purpose of this study is the detection of the association between MS and TNF-A gene promoter -376 polymorphism.In this study, 100 MS patients and 100 healthy controls were examined. Firstly, genomic DNA was extracted from human leukocyte nuclei isolated from whole blood by standard methods and then the promoter region of TNF-A including -376 polymorphism was amplified with PCR method. Secondly, the PCR products were digested with a digestion enzyme and genotype distributions and allele frequencies were determined. A strong association between MS and TNF-A -376 genotype distribution was detected        (p= 0,010). At the same time TNF-A -376 allele frequency was showed an association with MS (p=0,011), but this association indicates that GA genotype and A allele have protective effects. There is not an association between TNF-A -376 polymorphism and MS progression, but GA genotype and A allele may be the protective factors in the healthy people.

Anahtar Kelimeler

• Promoter,Polymorphism,Tumor Necrosis Factor Alpha (TNF-A),Multiple Sclerosis (MS)

Türk Popülasyonu’nda Tümor Nekroz Faktör Alfa Gen Polimorfizmi (-376) ile Multipl Skleroz Arasındaki İlişki

Öz

Bu araştırmanın amacı Multipl Skleroz (MS) ile Tümor Nekroz Faktör-Alfa (TNF-A) geninin promotör bölgesindeki (-376) gen polimorfizmi arasındaki ilişkininin ortaya çıkarılmasıdır. Yapılan çalışmada 100 MS hastası ve 100 sağlıklı kontrol olmak üzere toplam 200 birey incelenmiştir. Bunun için ilk olarak kandan DNA izolasyonu yapılmış ve elde edilen DNA’lardan TNF-A geninin promoter -376 bölgesi polimeraz zincir reaksiyonu yöntemi ile çoğaltılmıştır. İkinci aşamada ise elde edilen PCR ürünleri Tsp50 enzimi ile kesilerek genotip dağılımları ve allel frekansları incelenmiştir. Türk popülasyonunda MS hastalığı ile TNF-A    -376 polimorfizminin genotip dağılımı arasında kuvvetli bir ilişki bulunmuştur (p=0,010). Aynı zamanda TNF-A -376 polimorfizminin allel frekansı MS hastalığıyla bir ilişki göstermektedir (p=0.011). Bu ilişkide TNF-A -376 polimorfizminin heterozigot genotip G/A olduğu ve A allelinin koruyucu etkisi olduğu görülmüştür. TNF-A -376 polimorfizmi ile MS arasında hastalığın risk oluşturması açısından herhangi bir ilişki bulunmamıştır ancak G/A genotipi ve A allelinin sağlıklı kişilerde koruyucu etkisi olabildiği düşünülmektedir.  

Anahtar Kelimeler

• Promoter,Polimorfizm,Tümor Nekroz Faktör Alfa (TNF-A)

Kaynakça

1. Noseworthy JH., Lucchinetti C. ve Rodriguez M., Weinshenker BG. (2000). Multiple sclerosis. N Engl J Med., 343, 938-952.
2. Dyment DA., Sadovnick AD. ve Ebers GC. (1997). Genetics of multiple sclerosis. Hum Mol Genet, 6, 1693-1698.
3. Elahi MM., Asotra K., Matata BM. ve Mastana SS. (2009). Tumor necrosis factor alpha -308 gene locus promoter polymorphism: an analysis of association with health and disease. Biochim Biophys Acta, 1792, 163-172.
4. Zipp, F., Weber, F., Huber, S., Sotgiu, S., Czlonkowska, A., Holler, E., Albert, E., Weiss, E. H., Wekerle, H., ve Hohlfeld, R. (1995). Genetic control of multiple sclerosis: increased production of lymphotoxin and tumor necrosis factor-alpha by HLA-DR2+ T cells. Ann Neurol, 38, 723-730.
5. Hurwitz BJ. (2009).The diagnosis of multiple sclerosis and the clinical subtypes. Ann Indian Acad Neurol, 12, 226-230.
6. Kassiotis G. ve Kollias G. (2001). TNF and receptors in organ-specific autoimmune disease: multi-layered functioning mirrored in animal models. J Clin Invest, 107, 1507-1508.
7. Sharief MK.ve Hentges R. (1991). Association between tumor necrosis factor-alpha and disease progression in patients with multiple sclerosis. N Engl J Med, 325, 467-472.
8. Fargion S., Valenti L., Dongiovanni P. ve Fracanzani AL. (2004). TNFalpha promoter polymorphisms. Methods Mol Med, 98, 47-58.

9. Braun, N., Michel, U., Ernst, B. P., Metzner, R., Bitsch, A., Weber, F., ve Rieckmann, P. (1996). Gene polymorphism at position -308 of the tumor-necrosis-factor-alpha (TNF-alpha) in multiple sclerosis and it's influence on the regulation of TNF-alpha production. Neurosci Lett, 215(2), 75-78.
10. Hauser, S. L. (1995). Tumor necrosis factor: immunogenetics and disease. Ann Neurol, 38(5), 702-704.
11. Weinshenker, B. G., Wingerchuk, D. M., Liu, Q., Bissonet, A. S., Schaid, D. J., ve Sommer, S. S. (1997). Genetic variation in the tumor necrosis factor alpha gene and the outcome of multiple sclerosis. Neurology, 49(2), 378-385.
12. Wingerchuk, D., Liu, Q., Sobell, J., Sommer, S., ve Weinshenker, B. G. (1997). A population-based case-control study of the tumor necrosis factor alpha-308 polymorphism in multiple sclerosis. Neurology, 49(2), 626-628.
13. Bayley, J. P., de Rooij, H., van den Elsen, P. J., Huizinga, T. W., ve Verweij, C. L. (2001). Functional analysis of linker-scan mutants spanning the -376, -308, -244, and -238 polymorphic sites of the TNF-alpha promoter. Cytokine, 14(6), 316-323.
14. de Jong, B. A., Huizinga, T. W., Zanelli, E., Giphart, M. J., Bollen, E. L., Uitdehaag, B. M., Polman, C. H., ve Westendorp, R. G. (2002). Evidence for additional genetic risk indicators of relapse-onset MS within the HLA region. Neurology, 59(4), 549-555.
15. Wirz, S. A., Morale, M. C., Marchetti, B., Barr, A. M., Sotgiu, S., Rosati, G., Pugliatti, M., Sanna, M. V., Giliberto, O., Bartfai, T., ve Conti, B. (2004). High frequency of TNF alleles -238A and -376A in individuals from northern Sardinia. Cytokine, 26(4), 149-154.
16. Fernandez-Arquero, M., Arroyo, R., Rubio, A., Martin, C., Vigil, P., Conejero, L., Figueredo, M. A., ve de la Concha, E. G. (1999). Primary association of a TNF gene polymorphism with susceptibility to multiple sclerosis. Neurology, 53(6), 1361-1363.
17. Losonczi, E., Bencsik, K., Nagy, Z. F., Honti, V., Szalczer, E., Rajda, C., Illes, Z., Matyas, K., Rozsa, C., Csepany, T., Fuvesi, J., ve Vecsei, L. (2009). Tumour necrosis factor alpha gene (TNF-alpha) -376 polymorphism in Hungarian patients with primary progressive multiple sclerosis. J Neuroimmunol, 208(1-2), 115-118.
18. Drulovic, J., Popadic, D., Mesaros, S., Dujmovic, I., Cvetkovic, I., Miljkovic, D., Stojsavljevic, N., Pravica, V., Pekmezovic, T., Bogdanovic, G., Jarebinski, M., ve Mostarica Stojkovic, M. (2003). Decreased frequency of the tumor necrosis factor alpha -308 allele in Serbian patients with multiple sclerosis. Eur Neurol, 50(1), 25-29.
19. He, B., Navikas, V., Lundahl, J., Soderstrom, M., ve Hillert, J. (1995). Tumor necrosis factor alpha-308 alleles in multiple sclerosis and optic neuritis. J Neuroimmunol, 63(2), 143-147.
20. Huizinga, T. W., Westendorp, R. G., Bollen, E. L., Keijsers, V., Brinkman, B. M., Langermans, J. A., Breedveld, F. C., Verweij, C. L., van de Gaer, L., Dams, L., Crusius, J. B., Garcia-Gonzalez, A., van Oosten, B. W., Polman, C. H., ve Pena, A. S. (1997). TNF-alpha promoter polymorphisms, production and susceptibility to multiple sclerosis in different groups of patients. J Neuroimmunol, 72(2), 149-153.
21. Ristic, S., Lovrecic, L., Starcevic-Cizmarevic, N., Brajenovic-Milic, B., Sega Jazbec, S., Sepcic, J., Kapovic, M., ve Peterlin, B. (2007). Tumor necrosis factor-alpha-308 gene polymorphism in Croatian and Slovenian multiple sclerosis patients. Eur Neurol, 57(4), 203-207.
22. Maurer, M., Kruse, N., Giess, R., Kyriallis, K., Toyka, K. V., & Rieckmann, P. (1999). Gene polymorphism at position -308 of the tumor necrosis factor alpha promotor is not associated with disease progression in multiple sclerosis patients. J Neurol, 246(10), 949-954.
23. Lassmann, H., Bruck, W., ve Lucchinetti, C. F. (2007). The immunopathology of multiple sclerosis: an overview. Brain Pathol, 17(2), 210-218.
24. Knight, J. C., Udalova, I., Hill, A. V., Greenwood, B. M., Peshu, N., Marsh, K., ve Kwiatkowski, D. (1999). A polymorphism that affects OCT-1 binding to the TNF promoter region is associated with severe malaria. Nat Genet, 22(2), 145-150.