Interleukin-6 (-572) and Tumor Necrosis Factor α (-308 and -238) Polymorphisms and Risk of Knee Osteoarthritis in a Pakistani Population: A Case–Control Study

  • Syeda Kanwal Batool Naqvi
  • Maria Bibi
  • Iram MurtazaEmail author
  • Qamar JavedEmail author
Research paper
Part of the following topical collections:
  1. Biology


Osteoarthritis (OA) is one of the most common musculoskeletal diseases of adults worldwide. Inflammation plays a crucial part in OA pathology, but role of cytokines is still inconclusive. Purpose of this study was to assess the function of tumor necrosis factor α (TNF-α -308 and -238) and interleukin-6 (IL6-572) polymorphisms in OA disease susceptibility in a Pakistani population. 280 OA patients and 308 ethnically matched healthy controls were enrolled in the current study. Demographic data were collected from all participants via a questionnaire. Genomic DNA was isolated from all subjects. TNF-α (-308 G>A, -238 G>A) and IL6 (-572 G>C) polymorphisms in both groups were identified by PCR-coupled restriction fragment length polymorphism (PCR–RFLP) technique. GraphPad Prism software was used to perform statistical analysis. Genotypic and allelic frequencies were determined in both groups. Basic characteristics were mentioned as SD ± mean. p value above 0.05 was considered nonsignificant statistically. Age and body mass index (BMI) differences were not significant (> 0.05) between patients and control groups. Genotype frequencies were in agreement with Hardy–Weinberg equilibrium for all single nucleotide polymorphisms (SNPs) in control and patient group. TNF-α (-308, -238) GA+AA genotypes and IL6 (-572) GC+CC genotypes were considerably associated with higher risk of OA compared to homozygous wild-type genotypes (p < 0.01). Variant alleles were more expressed in knee OA patients as compared to healthy controls for all loci (p < 0.05). Our finding suggests there is an association between TNF-α -308G/A, -238G/A and IL6-572G/C polymorphisms and OA disease susceptibility in a Pakistani population. Further studies with large sample size and in diverse ethnic groups are vital to evaluate and confirm the function of these SNPs in OA disease pathology.


Knee osteoarthritis Single nucleotide polymorphism TNF-α IL6 Genotyping 



The authors are thankful to all participants of this study for their cooperation. We are especially grateful to Dr. Afrose Liaqat for providing valuable help in statistical analysis.

Compliance with Ethical Standards

Conflict of interest:

The authors declare that they have no conflict of interest.


  1. Abdel GSM, Ezzeldin N, Fawzy F, El-Boshym M (2017) The single-nucleotide polymorphism (SNP) of tumor necrosis factor α -308G/A gene is associated with early-onset primary knee osteoarthritis in an Egyptian female population. Clin Rheumatol 36(11):2525–2530CrossRefGoogle Scholar
  2. Altman R, Asch E, Bloch D, Bole G, Borenstein D, Brandt K et al (1986) Development of criteria for the classification and reporting of osteoarthritis, classification of osteoarthritis of the knee. Diagnostic and Therapeutic Criteria Committee of the American Rheumatism Association. Arthritis Rheumatol 29:1039–1049CrossRefGoogle Scholar
  3. Bin J, Shi J, Cheng X, Zhou J, Zhou Q, Cao C (2013) Association analysis of two candidate polymorphisms in the Tumor Necrosis Factor-α gene with osteoarthritis in a Chinese population. Int Orthop 37(10):2061–2063CrossRefGoogle Scholar
  4. Brinkman BM, Zuijdeest D, Kaijzel EL, Breedveld FC, Verweij CL (1996) Relevance of the tumor necrosis factor alpha (TNF alpha) -308 promoter polymorphism in TNF alpha gene regulation. J Inflamm 46(1):32–41Google Scholar
  5. Chen D, Shen J, Zhao W, Wang T, Han L, Hamilton JL et al (2017) Osteoarthritis: toward a comprehensive understanding of pathological mechanism. Bone Res 5:16044CrossRefGoogle Scholar
  6. Fernandes JC, Martel-Pelletier J, Pelletier JP (2002) The role of cytokines in osteoarthritis pathophysiology. Biorheology 39:237–246Google Scholar
  7. Fernandes MTP, Fernandes KBP, Marquez AS, Cólus MS, Souza MF, Santos JPM et al (2015) Association of interleukin-6 gene polymorphism (rs1800796) with severity and functional status of osteoarthritis in elderly individuals. Cytokine 75(2):316–320CrossRefGoogle Scholar
  8. Han L, Song JH, Yoon JH, Paer YG, Lee SW, Choi YJ et al (2012) TNF-a and TNF-β polymorphisms are associated with susceptibility to osteoarthritis in a Korean population. Korean J Pathol 46(1):30–37CrossRefGoogle Scholar
  9. Haq SA (2011) Osteoarthritis of the knees in the COPCORD world. Int J Rheum Dis 14:122–129CrossRefGoogle Scholar
  10. Helmy N, Maly FE, Bestmann L et al (2001) Detection of the single-base substitution -174 G > C in the interleukin-6 gene by real-time polymerase chain reaction: comment on the article by Moos et al. Arthritis Rheumatol 44:2213–2214CrossRefGoogle Scholar
  11. Jamil K, Archana J, Javeed A, Sindhu J, Sk Yerra (2017) TNF-alpha -308G/A and -238G/A polymorphisms and its protein network associated with type 2 diabetes mellitus. Saudi J Biol Sci 24(6):1195–1203CrossRefGoogle Scholar
  12. Jordanies N, Eskdale J, Stuart R, Gallagher G (2000) Allele associations reveal four prominent haplotypes at the human interleukin-6 (IL-6) locus. Genes Immun 1:451–455CrossRefGoogle Scholar
  13. Kämäräinen OP, Solovieva S, Vehmas T, Luoma K, Riihimäki H, Ala-Kokko L et al (2008) Common interleukin-6 promoter variants associate with the more severe forms of distal interphalangeal osteoarthritis. Arthritis Res Ther 10(1):R21. CrossRefGoogle Scholar
  14. Kaneko S, Satoh T, Chiba J, Ju C, Inoue K, Kagawa J (2000) Interleukin-6 and interleukin-8 levels in serum and synovial fluid of patients with osteoarthritis. Cytokines Cell Mol Ther 6(2):71–79CrossRefGoogle Scholar
  15. Kou S, Wu Y (2014) Meta-analysis of tumor necrosis factor alpha -308 polymorphism and knee osteoarthritis risk. BMC Musculoskeletal Disord 15:373CrossRefGoogle Scholar
  16. Lee JY, Dong SM, Kim SY, Yoo NJ, Lee SH, Park WS (1998) A simple, precise and economical micro dissection technique for analysis of genomic DNA from archival tissue sections. Virchows Arch 433(4):305–309CrossRefGoogle Scholar
  17. Lee YH, Ji JD, Bae SC, Song GG (2010) Associations between tumor necrosis factor-alpha (TNF-alpha) -308 and -238 G/A polymorphisms and shared epitope status and responsiveness to TNF-alpha blockers in rheumatoid arthritis: a meta-analysis update. J Rheumatol 37(4):740–746CrossRefGoogle Scholar
  18. Liaquat A, Asifa GZ, Zeenat A, Javed Q (2014) Polymorphisms of tumor necrosis factor-alpha and interleukin-6 gene and C-reactive protein profiles in patients with idiopathic dilated cardiomyopathy. Ann Saudi Med 34(5):407–414CrossRefGoogle Scholar
  19. Liaquat A, Shauket U, Ahmad W, Javed Q (2015) The tumor necrosis factor-α-238G/A and IL-6 -572G/C gene polymorphisms and the risk of idiopathic dilated cardiomyopathy: a meta-analysis of 25 studies including 9493 cases and 13,971 controls. Clin Chem Lab Med 53:307–318Google Scholar
  20. Lieberthal J, Sambamurthy N, Scanzello CR (2015) Inflammation in joint injury and post-traumatic osteoarthritis. Osteoarthr Cartil 23(11):1825–1834CrossRefGoogle Scholar
  21. Livshits G, Zhai G, Hart DJ, Kato BS, Wang H, Williams FM et al (2009) Interleukin-6 is a significant predictor of radiographic knee osteoarthritis: the Chingford study. Arthritis Rheumatol 60(7):2037–2045CrossRefGoogle Scholar
  22. Manolagas SC, Jilka RL (1995) Bone Marrow, Cytokines, and Bone Remodeling — Emerging Insights into the Pathophysiology of Osteoporosis. N Engl J Med 332(5):305–311CrossRefGoogle Scholar
  23. Marinou I, Healy J, Mewar D, Moore DJ, Dickson MC, Binks MH et al (2007) Association of interleukin-6 and interleukin-10 genotypes with radiographic damage in rheumatoid arthritis is dependent on autoantibody status. Arthritis Rheumatol 56(8):2549–2556CrossRefGoogle Scholar
  24. Munoz VJF, Oregon RE, Rangel VH, Martinez BGE, Castaneda SE, Salgado GL et al (2014) High expression of TNF alpha is associated with -308 and -238 TNF alpha polymorphisms in knee osteoarthritis. Clin Exp Med 14:61–67CrossRefGoogle Scholar
  25. Orita S, Koshi T, Mitsuka T, Miyagi M, Inoue G, Arai G et al (2011) Associations between proinflammatory cytokines in the synovial fluid and radiographic grading and pain-related scores in 47 consecutive patients with osteoarthritis of the knee. BMC Musculoskelet Disord 12(1):144CrossRefGoogle Scholar
  26. Osiri M, McNicholl J, Moreland LW, Bridges SL (1999) A novel single nucleotide polymorphism and five probable haplotypes in the 5′ flanking region of the IL-6 gene in African-Americans. Genes Immun 1(2):166–167CrossRefGoogle Scholar
  27. Özler K, Aktaş E, Atay Ç, Yılmaz B, Arıkan M, Güngör Ş (2016) Serum and knee synovial fluid matrixmetalloproteinase-13 and tumor necrosis factor-alpha levels in patients with late stage osteoarthritis. Acta Orthop Traumatol Turc 50(6):670–673CrossRefGoogle Scholar
  28. Pelletier JP, Martel-Pelletier J, Abramson SB (2001) Osteoarthritis, an inflammatory disease: potential implication for the selection of the new therapeutic targets. Arthritis Rheumatol 44(6):1237–1247CrossRefGoogle Scholar
  29. Sezgin M, Barlas IO, Ankarali HC, Altintas ZM, Türkmen E, Gökdoğan (2008) Tumor necrosis factor alpha -308G/A gene polymorphism: lack of association with knee osteoarthritis in a Turkish population. Clin Exp Rheumatol 5:763–768Google Scholar
  30. Shlopov BV, Gumanovskaya ML, Hasty KA (2000) Autocrine regulation of collagenase 3 (matrix metalloproteinase 13) during osteoarthritis. Arthritis Rheumatol 43:195–205CrossRefGoogle Scholar
  31. Sobhan MR, Mahdinezhad YM, Aghili K, Zare SM et al (2018) Association of TNF-α-308 G > A and -238G > A polymorphisms with knee osteoarthritis risk: a case-control study and meta-analysis. J Orthop 15(3):747–753CrossRefGoogle Scholar
  32. Sousa E, Caetano LJ, Pint P, Pimentel F, Teles J, Canhã H et al (2009) Ankylosing spondylitis susceptibility and severity–contribution of TNF gene promoter polymorphisms at positions -238 and -308. Ann N Y Acad Sci 1173:581–588CrossRefGoogle Scholar
  33. Valdes AM, Arden NK, Tamm A, Kisand K, Doherty S, Pola E et al (2010) A meta-analysis of interleukin-6 promoter polymorphisms on risk of hip and knee osteoarthritis. Osteoarthr Cartil 18(5):699–704CrossRefGoogle Scholar
  34. Wang J, Platt A, Upmanyu R, Germer S et al (2013) IL-6 pathway-driven investigation of response to IL-6 receptor inhibition in rheumatoid arthritis. BMJ Open 3:e003199CrossRefGoogle Scholar

Copyright information

© Shiraz University 2019

Authors and Affiliations

  1. 1.Department of Biochemistry, Faculty of Biological SciencesQuaid-i-Azam UniversityIslamabadPakistan
  2. 2.Preston Institute of Nano Science and TechnologyPreston UniversityIslamabadPakistan

Personalised recommendations