Molecular Biology Reports

, Volume 46, Issue 2, pp 2131–2138 | Cite as

Association of rs6259 polymorphism with SHBG levels and Poly Cystic Ovary Syndrome in Indian population: a case control study

  • Richa Bhatnager
  • Alka Senwal
  • Smiti Nanda
  • Amita S. DangEmail author
Original Article


Polycystic ovary syndrome (PCOS) is the most common endocrinopathy of reproductive-aged women. PCOS reflects a number of possible etiologies but its pathophysiology is still unclear. The principal abnormality of the syndrome is hyperandrogenism (70–80%). The access of androgens to target tissues is regulated by sex hormone-binding globulin (SHBG), a transport protein secreted by liver i.e. specific for androgens. Present study was done to find the association of rs6259 polymorphism with SHBG levels and Poly Cystic Ovary Syndrome in Indian population. Present study was a case control study. 400 subjects were enrolled for the study and serum SHBG levels and D327N polymorphism were measured. The D327N polymorphism (wild-type and variant allele) was detected using PCR-RFLP method (restriction enzyme Bbs-I). PCOS group was found to have significantly lower SHBG levels than healthy controls. There was no significant difference in genotype distribution between PCOS and controls (χ2 = 1.0335, p = 0.59). Significant difference in SHBG levels of PCOS and control group highlights the potential of SHBG as a biomarker for PCOS. However, no significant difference in genotype distribution between PCOS and controls provided an insight that rs6259 polymorphism is not associated with the risk of PCOS and SHBG levels.


PCOS SHBG Diagnostic marker Susceptibility marker Polymorphism 



Authors acknowledge DST-INSPIRE division and DBT-HRD to support the research work and Centre for Medical biotechnology, MDU, Rohtak, Haryana, India to provide a research platform to carry out research work.

Compliance with ethical standards

Conflict of interest

Authors declare no conflict of interest.

Supplementary material

11033_2019_4665_MOESM1_ESM.doc (224 kb)
Supplementary material 1 Fig. 1 Study Population segregation: study population was screened according to Rotterdam criteria. SHBG levels measurement and PCR-RFLP pattern was analyzed; Subjects were also segregated on BMI to find its association with polymorphism under study. PCOS patients were segregated on the basis of presence and absence of Hyperandrogenism (HA) to evaluate the role of rs6259 in predisposition of hyperandrogenism as well. Fig. 2 Receiver’s Operative curve for SHBG as a diagnostic marker for PCOS: Significant difference in PCOS (Poly Cystic Ovary Syndrome) and control population highlights the potential of SHBG (Sex hormone binding globulin) to be used as a diagnostic marker for the identification of the syndrome ROC curve with a specificity and sensitivity 93.51 and 99.38% respectively. Fig. 3 Receiver’s Operative curve for SHBG as a susceptibility marker for PCOS: Reduced SHBG (Sex hormone binding globulin) levels were found in obese PCOS (Poly Cystic Ovary Syndrome) patients than lean PCOS patients. Hence it can be predicted as a marker to differentiate obese and lean population with a specificity and sensitivity score 90.57 and 98.67% respectively. Fig. 4 Receiver’s Operative curve for SHBG as a phenotypic differential marker for PCOS: SHBG (Sex hormone binding globulin) levels were low in Hyperandrogenic PCOS patients than Non hyperandrogenic PCOS (Poly Cystic Ovary Syndrome) subjects. The result can be used as a marker with a specificity and sensitivity score 94.69 and 97.31% respectively. Supplementary Fig. 5 Restriction analysis of PCR product on 3% agarose gel: Lane 1: 50 bp ladder, lane 2: AA genotype (290 bp), Lane3 and 5: GG genotype (223 bp product and 67 bp product: not visible), lane 4: GA genotype (290,223, 67 bp digested product) (DOC 224 KB)


  1. 1.
    Pugeat M, Crave JC, Tourniaire J et al (1996) Clinical utility of sex hormone-binding globulin measurement. Horm Res Paediatr 45:148–155CrossRefGoogle Scholar
  2. 2.
    Anderson DC (1974) Sex-hormone-binding globulin. Clin Endocrinol 3:69–96CrossRefGoogle Scholar
  3. 3.
    Rosner W (1991) Plasma steroid-binding proteins. Endocrinol Metab Clin North Am 20:697–720CrossRefPubMedGoogle Scholar
  4. 4.
    Dunaif A, Segal KR, Futterweit W et al (1989) Profound peripheral insulin resistance, independent of obesity, in polycystic ovary syndrome. Diabetes 38:1165–1174CrossRefPubMedGoogle Scholar
  5. 5.
    Bozdag G, Mumusoglu S, Zengin D et al (2016) The prevalence and phenotypic features of polycystic ovary syndrome: a systematic review and meta-analysis. Hum Reprod 31:2841–2855CrossRefPubMedGoogle Scholar
  6. 6.
    Rotterdam ES, ASRM-Sponsored PCOS Consensus Workshop Group (2004) Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome (PCOS). Hum Reprod 19(1):41CrossRefGoogle Scholar
  7. 7.
    Alexiou E, Hatziagelaki E, Pergialiotis V et al (2017) Hyperandrogenemia in women with polycystic ovary syndrome: prevalence, characteristics and association with body mass index. Horm Mol Biol Clin Investig 29:105–111PubMedGoogle Scholar
  8. 8.
    Kalyani RR, Franco M, Dobs AS et al (2009) The association of endogenous sex hormones, adiposity, and insulin resistance with incident diabetes in postmenopausal women. J Clin Endocrinol Metab 94:4127–4135CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Le TN, Nestler JE, Strauss JF et al (2012) Sex hormone-binding globulin and type 2 diabetes mellitus. Trends Endocrinol Metab 23:32–40CrossRefPubMedGoogle Scholar
  10. 10.
    Zha XY, Hu Y, Pang XN et al (2016) The association between sex hormone-binding globulin gene polymorphism with bone mineral density. Steroids 106:9–18CrossRefPubMedGoogle Scholar
  11. 11.
    El Tarhouny SA, Zakaria SS, Abdu-Allah AM et al (2015) Study of sex hormone-binding globulin gene polymorphism and risk of type 2 diabetes mellitus in Egyptian men. West Indian Med J 64:338PubMedGoogle Scholar
  12. 12.
    .Zhang LS, Yuan F, Guan X et al (2014) Association of genetic polymorphisms in HSD17B1, HSD17B2 and SHBG genes with hepatocellular carcinoma risk. Pathol Oncol Res 20:661–666CrossRefPubMedGoogle Scholar
  13. 13.
    Xu WH, Zheng W, Cai Q et al (2008) The Asp 327 Asn polymorphism in the sex hormone-binding globulin gene modifies the association of soy food and tea intake with endometrial cancer risk. Nutr Cancer 60:736–743CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Safarinejad MR, Shafiei N, Safarinejad S (2011) Association of the (TAAAA) n repeat and Asp327Asn polymorphisms in the sex hormone-binding globulin (SHBG) gene with idiopathic male infertility and relation to serum SHBG concentrations. J Steroid Biochem Mol Biol 123:37–45CrossRefPubMedGoogle Scholar
  15. 15.
    Bendlová B, Zavadilová J, Vaňková M et al (2007) Role of D327N sex hormone-binding globulin gene polymorphism in the pathogenesis of polycystic ovary syndrome. J Steroid Biochem Mol Biol 104:68–74CrossRefPubMedGoogle Scholar
  16. 16.
    Martínez-García M, Gambineri A, Alpañés M et al (2012) Common variants in the sex hormone-binding globulin gene (SHBG) and polycystic ovary syndrome (PCOS) in Mediterranean women. Hum Reprod 27:3569–3576CrossRefPubMedGoogle Scholar
  17. 17.
    Wickham EP III, Ewens KG, Legro RS et al (2011) Polymorphisms in the SHBG gene influence serum SHBG levels in women with polycystic ovary syndrome. J Clin Endocrinol Metab 96:E719–E727CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Hacıhanefioğlu B, Aybey B, Hakan Özön Y et al (2013) Association of anthropometric, androgenic and insulin-related features with polymorphisms in exon 8 of SHBG gene in women with polycystic ovary syndrome. Gynecol Endocrinol 29:361–364CrossRefPubMedGoogle Scholar
  19. 19.
    Thaler MA, Seifert-Klauss V, Luppa PB (2015) The biomarker sex hormone-binding globulin—from established applications to emerging trends in clinical medicine. Best Pract Res Clin Endocrinol Metab 29:749–760CrossRefPubMedGoogle Scholar
  20. 20.
    Toljan K, Grgić F, Pavičić Baldani D et al (2016) Sex hormone binding globulin (SHBG) as a marker of clinical disorders. Coll Antropol 40:199–209Google Scholar
  21. 21.
    Fabbri E, An Y, Gonzalez-Freire M et al (2016) Bioavailable testosterone linearly declines over a wide age spectrum in men and women from the Baltimore longitudinal study of aging. J Gerontol A Biol Sci Med Sci 71:1202–1209CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Azrad M, Gower BA, Hunter GR et al (2012) Intra-abdominal adipose tissue is independently associated with sex-hormone binding globulin in premenopausal women. Obesity 20:1012–1015CrossRefPubMedGoogle Scholar
  23. 23.
    Kim JS, Kang HT, Shim JY (2012) The association between the triglyceride to high-density lipoprotein cholesterol ratio with insulin resistance (HOMA-IR) in the general Korean population: based on the National Health and Nutrition Examination Survey in 2007–2009. Diabetes Res Clin Pract 97:132–138CrossRefPubMedGoogle Scholar
  24. 24.
    Azziz R, Carmina E, Dewailly D et al (2009) The Androgen Excess and PCOS Society criteria for the polycystic ovary syndrome: the complete task force report. Fertil Steril 91:456–488CrossRefPubMedGoogle Scholar
  25. 25.
    Abu-Hijleh TM, Gammoh E, Al-Busaidi AS et al (2016) Common variants in the sex hormone-binding globulin (SHBG) gene influence SHBG levels in women with polycystic ovary syndrome. Ann Nutr Metab 68:66–74CrossRefPubMedGoogle Scholar
  26. 26.
    White MJ, Eren F, Agirbasli D et al (2015) SHBG gene polymorphism (rs1799941) associates with metabolic syndrome in children and adolescents. PLoS ONE 10:e0116915CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Baldani DP, Skrgatic L, Cerne JZ et al (2015) Association between serum levels and pentanucleotide polymorphism in the sex hormone binding globulin gene and cardiovascular risk factors in females with polycystic ovary syndrome. Mol Med Rep 11:3941–3947CrossRefPubMedGoogle Scholar
  28. 28.
    Markatseli AE, Hatzi E, Pamporaki C et al (2014) Association of the (TAAAA) n repeat polymorphism of SHBG gene with the age at menopause in Greek postmenopausal women. Maturitas 78:113–116CrossRefPubMedGoogle Scholar
  29. 29.
    Svartberg J, Schirmer H, Wilsgaard T et al (2014) Single-nucleotide polymorphism, rs1799941 in the sex hormone-binding globulin (SHBG) gene, related to both serum testosterone and SHBG levels and the risk of myocardial infarction, type 2 diabetes, cancer and mortality in men: the Tromsø study. Andrology 2:212–218CrossRefPubMedGoogle Scholar
  30. 30.
    Sunbul M, Eren F, Nacar C et al (2013) Sex hormone binding globulin gene polymorphisms and metabolic syndrome in postmenopausal Turkish women. Cardiol J 20:287–293CrossRefPubMedGoogle Scholar
  31. 31.
    Monteiro C, Sousa MV, Ribeiro R et al (2013) Genetic variants in AR and SHBG and resistance to hormonal castration in prostate cancer. Med Oncol 30:490CrossRefPubMedGoogle Scholar
  32. 32.
    Saltiki K, Stamatelopoulos K, Voidonikola P et al (2011) Association of the SHBG gene promoter polymorphism with early markers of atherosclerosis in apparently healthy women. Atherosclerosis 219:205–210CrossRefPubMedGoogle Scholar
  33. 33.
    Vanková M, Prazáková S, Lukásová P et al (2010) Extremely low SHBG level in consequence of Pro156Leu SHBG polymorphism—case reports of two women with polycystic ovary syndrome. Vnitrni Lekarstvi 56:1292–1295PubMedGoogle Scholar
  34. 34.
    Xita N, Milionis HJ, Galidi A et al (2011) The (TAAAA) n polymorphism of the SHBG gene in men with the metabolic syndrome. Exp Clin Endocrinol Diabetes 119:126–128CrossRefPubMedGoogle Scholar
  35. 35.
    Piotrowski P, Gasik R, Lianeri M et al (2010) Asp327Asn polymorphism of sex hormone-binding globulin gene is associated with systemic lupus erythematosus incidence. Mol Biol Rep 37:235–239CrossRefPubMedGoogle Scholar
  36. 36.
    Cousin P, Calemard-Michel L, Lejeune H et al (2004) Influence of SHBG gene pentanucleotide TAAAA repeat and D327N polymorphism on serum sex hormone-binding globulin concentration in hirsute women. J Clin Endocrinol Metab 89:917–924CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Centre for Medical BiotechnologyMaharshi Dayanand UniversityRohtakIndia
  2. 2.Pandit Bhagwat Dayal Sharma Institute of Medical SciencesRohtakIndia

Personalised recommendations