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Estrogen receptor alpha gene analysis in osteoporosis and familial osteoporosis

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Abstract

Estrogens are important determinants of bone mineral density (BMD) mediating their effects via estrogen receptor α (ERα) and β (ERβ). The strong genetic predisposition to osteoporosis, and the fact that alterations in the aminoterminal region of ERα have been linked to bone disturbances, prompted us to identify genetic alterations in exon 1 and exon 2 of ERα in osteoporotic individuals. Sixty-two unrelated normal subjects (age 46.1±9.5 years) and 72 unrelated osteoporotic subjects (age 52.3±7.9 years) were studied. Their menopausal status was pre- and perimenopausal. We also included 30 related osteoporotic individuals (mother-daughter or sister-sister relationship) (age 46.2±12.8 years) belonging to 14 families who where also pre- and perimenopausal. DNA was extracted from peripheral blood, exons 1 and 2 were amplified by polymerase chain reaction (PCR) and were further submitted to denaturing gradient gel electrophoresis (DGGE), single stranded conformational polymorphism (SSCP), restriction fragment length polymorphism (RFLP) and sequence analysis. Bone turnover markers were also determined. Two polymorphisms were identified in exon 1 (codons 10 and 87) in both normal and osteoporotic women. Statistical analysis revealed no difference (P>0.05) in the ERα genotype frequencies within osteoporotic families as compared with the same genotypes in the unrelated normal or osteoporotic subjects. Codon 10, codon 87 polymorphisms were not related to BMD or bone turnover markers. No other mutations were found in exons 1 and 2 in all subjects studied. Genetic alterations in exons 1 and 2 of ERα are not associated to osteoporosis and familial osteoporosis. Moreover, the codon 10 and codon 87 polymorphisms do not seem to be correlated with BMD and bone turnover markers.

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References

  1. Rosen CJ, Kessenich CR (1997) The pathophysiology and treatment of postmenopausal osteoporosis. An evidence-based approach to estrogen replacement therapy. Endocr Metabol Clin N Am 26:295–311

    CAS  Google Scholar 

  2. Manolagas SC, Kousteni S, Jilka RL (2002) Sex steroids and bone. Recent Prog Horm Res 57:385–409

    Article  CAS  PubMed  Google Scholar 

  3. Eisman JA (1999) Genetics of osteoporosis. Endocr Rev 20:788–804

    Article  CAS  PubMed  Google Scholar 

  4. Seeman E, Hopper JL, Bach LA, Cooper ME, Parkinson E, McKay J, Jerums G (1989) Reduced bone mass in daughters of women with osteoporosis. N Engl J Med 320:554–558

    CAS  PubMed  Google Scholar 

  5. Harris M, Nguyen TV, Howard GM, Kelly PJ, Eisman JA (1998) Genetic and environmental correlations between bone formation and bone mineral density: a twin study. Bone 22:141–145

    Article  CAS  PubMed  Google Scholar 

  6. Kobayashi S, Inoue S, Hosoi T, Ouchi Y, Shiraki M, Orimo H (1996) Association of bone mineral density with polymorphism of the estrogen receptor gene. J Bone Miner Res 11:306–311

    CAS  PubMed  Google Scholar 

  7. Sano M, Inoue S, Hosoi T, Ouchi Y, Emi M, Shiraki M, Orimo H (1995) Association of estrogen receptor dinucleotide repeat polymorphism with osteoporosis. Biochem Biophys Res Commun 217:378–383

    Article  CAS  PubMed  Google Scholar 

  8. Han OK, Moon IG, Kang YS, Chung HY, Min HK, Han IK (1997) Nonassociation of estrogen receptor genotypes with bone mineral density and estrogen responsiveness to hormone replacement therapy in Korean postmenopausal women. J Clin Endocrinol Metab 82:991–995

    Article  CAS  PubMed  Google Scholar 

  9. Mizunuma H, Hosoi T, Okano H, Soda M, Tokizawa T, Kagami I, Miyamoto S, Ibuki Y, Inoue S, Shiraki M, Ouchi Y (1997) Estrogen receptor gene polymorphism and bone mineral density at the lumbar spine of pre- and postmenopausal women. Bone 21:379–383

    Article  CAS  PubMed  Google Scholar 

  10. Langdahl BL, Lokke E, Carstens M, Stenkjaer LL, Eriksen EF (2000) A TA repeat polymorphism in the estrogen receptor gene is associated with osteoporotic fractures but polymorphisms in the first exon and intron are not. J Bone Miner Res 15:2222–2230

    CAS  PubMed  Google Scholar 

  11. Liu YZ, Liu YJ, Recker RR, Deng HW (2003) Molecular studies of identification of genes for osteoporosis: the 2002 update. J Endocrinol 177:147–196

    CAS  PubMed  Google Scholar 

  12. Albagha OM, McGuigan FE, Reid DM, Ralston SH (2001) Estrogen receptor alpha gene polymorphisms and bone mineral density: haplotype analysis in women from the United Kingdom. J Bone Miner Res 16:128–134

    CAS  PubMed  Google Scholar 

  13. Becherini L, Gennari L, Masi L, Mansani R, Massart F, Morelli A, Falchetti A, Gonnelli S, Fiorelli G, Tanini A, Brandi ML (2000) Evidence of a linkage disequilibrium between polymorphisms in the human estrogen receptor α gene and their relationship to bone mass variation in postmenopausal Italian women. Hum Mol Genet 9:2043–2050

    Article  CAS  PubMed  Google Scholar 

  14. Ioannidis JPA, Stavrou I, Trikalinos TA, Zois C, Brandi ML, Gennari L, Albagha O, Ralston SH, Tsatsoulis A (2002) Association of polymorphisms of the estrogen receptor α gene with bone mineral density and fracture risk in women: a meta-analysis. J Bone Miner Res 17:2048–2060

    CAS  PubMed  Google Scholar 

  15. Tsukamoto K, Yoshida H, Watanabe S, Suzuki T, Miyao M, Hosoi T, Orimo H, Emi M (1999) Association of radial bone mineral density with CA repeat polymorphism at the interleukin 6 locus in postmenopausal Japanese women. J Hum Genet 44:148–151

    Article  CAS  PubMed  Google Scholar 

  16. Green S, Walter P, Kuma V, Krust A, Bornert J, Argos P, Chambon P (1986) Human oestrogen receptor cDNA, sequence, expression and homology to v-erb-A. Nature 320:134–139

    CAS  PubMed  Google Scholar 

  17. Mosselman S, Polman J, Dijkema R (1996) ERβ: identification and characterization of a novel human estrogen receptor. FEBS Lett 392:49–53

    Article  CAS  PubMed  Google Scholar 

  18. Beato M, Herrlich P, Schütz G (1995) Steroid hormone receptors: many actors in search of a plot. Cell 83:851–857

    Article  CAS  PubMed  Google Scholar 

  19. Srivastava S, Weitzmann MN, Cenci S, Ross FP, Adler S, Pacifici R (1999) Estrogen decreases TNF gene expression by blocking JNK activity and the resulting production of C-Jun and Jun D. J Clin Invest 104:503–513

    CAS  PubMed  Google Scholar 

  20. Quaedackers ME, Van Den Brink CE, Wissink S, Schreurs RHMM, Gustafsson JA, Van Der Saag PT, Van Der Burg B (2001) 4-Hydroxytamoxifen trans-represses nuclear factor-kappa B activity in human osteoblastic U2-OS cells through estrogen receptor (ER) alpha, and not through ER beta. Endocrinology 142:1156–1166

    Article  CAS  PubMed  Google Scholar 

  21. American Society for Reproductive Medicine (2001) The menopausal transition. Fertil Steril 76:875–879

    Google Scholar 

  22. Kassi EN, Vlachoyiannopoulos PG, Moutsopoulos HM, Sekeris CE, Moutsatsou P (2001) Molecular analysis of estrogen receptor alpha and beta in lupus patients. Eur J Clin Invest 31:86–93

    Article  CAS  PubMed  Google Scholar 

  23. Puchner AM, Tziotis J, Evangelopoulos D, Fountas L, Vlahos G, Creatsas G, Sekeris CE, Moutsatsou P (2001) Estrogen receptor alpha gene analysis in preeclampsia. Steroids 66:695–700

    Article  PubMed  Google Scholar 

  24. Lerman LS, Silverstein K (1987) Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Meth Enzymol 155:482–501

    Article  CAS  PubMed  Google Scholar 

  25. Moutsatsou P, Fountas L, Coulocheri S, Kassi E, Sekeris CE (1999) The oestrogen receptor codon 10 polymorphism detected in breast cancer is also present in non-malignant cells. J Cancer Res Clin Oncol 125:214–218

    Article  CAS  PubMed  Google Scholar 

  26. Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249

    Google Scholar 

  27. Ogawa S, Hosoi T, Shiraki M, Orimo H, Emi M, Muramatsu M, Inoue S (2000) Association of estrogen receptor beta gene polymorphism with mineral density. Biochem Biophys Res Commun 269: 537–541

    Article  CAS  PubMed  Google Scholar 

  28. Smith EP, Boyd J, Frank GR, Takahashi H, Cohen RM, Specker B, Williams TC, Lubahn DB, Korach KS (1994) Estrogen resistance caused by a mutation in the estrogen-receptor gene in a man. N Engl J Med 331:1056–1061

    Article  CAS  PubMed  Google Scholar 

  29. Lehrer S, Rabin J, Kalir T, Schacter BS (1993) Estrogen receptor variant and hypertension in women. Hypertension 21:439–441

    CAS  PubMed  Google Scholar 

  30. Deng HW, Li J, Li JL, Johnson M, Gong G, Recker RR (1999) Association of VDR and estrogen receptor genotypes with bone mass in postmenopausal Caucasian women: different conclusions with different analyses and the implications. Osteoporos Int 9:499–507

    CAS  PubMed  Google Scholar 

  31. Hernandez CJ, Beaupre GS, Carter DR (2003) A theoretical analysis of the relative influences of peak BMD, age-related bone loss and menopause on the development of osteoporosis. Osteoporos Int [Epub ahead of print] 14:843–847

    Google Scholar 

  32. Bagger YZ, Jorgensen HL, Heegaard AM, Bayer L, Hansen L (2000) No major effect of estrogen receptor gene polymorphisms of mineral density or bone loss in postmenopausal Danish women. Bone 26:111–116

    Article  CAS  PubMed  Google Scholar 

  33. Turner RT, Rigs BL, Spelsberg TC (1994) Skeletal effects of estrogen. Endocr Rev 15:275–300

    Article  CAS  PubMed  Google Scholar 

  34. Lewin B (1997) Genes, 6th edn. Oxford University Press, Oxford, p 700

  35. Garcia T, Lehrer S, Bloomer WD, Schachter B (1988) A variant estrogen receptor messenger ribonucleic acid is associated with reduced level of estrogen binding in human mammary tumors. Mol Endocrinol 2:785–791

    CAS  PubMed  Google Scholar 

  36. Lehrer S, Sanchez M, Song HK, Dalton J, Levine E, Savoretti P, Thung SN, Schachter B (1990) Oestrogen receptor B-region polymorphism and spontaneous abortion in women with breast cancer. Lancet 17:622–624

    Article  Google Scholar 

  37. Roodi N, Bailey LR, Kao WY, Verrier CS, Yee CJ, Dupont WD, Parl FF (1995) Estrogen receptor gene analysis in estrogen receptor-positive and receptor-negative primary breast cancer. J Natl Cancer Inst 8715:446–451

    Google Scholar 

  38. Fujimmoto J, Hirose R, Ichigo S, Sakaguchi H, Tamaya T (1998) DNA polymorphism in B-domain of the estrogen receptor-alpha among Japanese women. Steroids 63:146–148

    Article  PubMed  Google Scholar 

  39. Couse JF, Korach KS (1999) Estrogen receptor null mice: what have we learned and where will they lead us? Endocr Rev 20:358–417

    Article  CAS  PubMed  Google Scholar 

  40. Windahl SH, Andersson G, Gustafsson JA (2002) Elucidation of estrogen receptor function in bone with the use of mouse models. Trends Endocrinol Metab 13:195

    Article  CAS  PubMed  Google Scholar 

  41. Denger S, Reid G, Kos M, Flouriot G, Parsch D, Brand H, Korach KS, Sonntag-Buck V, Gannon F (2001) ERalpha gene expression in human primary osteoblasts: evidence for the expression of two receptor proteins. Mol Endocrinol 15:2064–2077

    Article  CAS  PubMed  Google Scholar 

  42. Rosenbrock H, Seifert-Klauss V, Kaspar S, Busch R, Luppa PB (2002) Changes of biochemical bone markers during the menopausal transition. Clin Chem Lab Med 40:143–151

    CAS  PubMed  Google Scholar 

  43. Seifert-Klauss V, Mueller JE, Luppa P, Probst R, Wilker J, Hoss C, Treumann T, Kastner C, Ulm K (2002) Bone metabolism during the perimenopausal transition: a prospective study. Maturitas 41:23–33

    Article  CAS  PubMed  Google Scholar 

  44. Sheehan D, Bennett T, Cashman KD (2001) An assessment of genetic markers as predictors of bone turnover in healthy adults. J Endocrinol Invest 24:236–245

    CAS  PubMed  Google Scholar 

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Acknowledgements

This study was supported by the General Secretariat for Research and Technology of the Ministry of Development, Greece (PAVE research program) in cooperation with the Diagnostic Center “BIOMED SA”.

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Authors and Affiliations

  1. Department of Biological Chemistry, Medical School, National University of Athens, 75 Mikras Asias Street Goudi, 115 27, Athens, Greece

    L. Fountas, C. E. Sekeris, E. Kassi & P. Moutsatsou

  2. Department of Pathophysiology, Medical School, National University of Athens, 75 Mikras Asias street Goudi, 115 27, Athens, Greece

    M. Anapliotou

  3. Department of Animal Breeding, Agriculture University of Athens, Iera Odos 75, GR 118 55, Athens, Greece

    A. Kominakis

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  1. L. Fountas
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  2. M. Anapliotou
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Correspondence to P. Moutsatsou.

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Fountas, L., Anapliotou, M., Kominakis, A. et al. Estrogen receptor alpha gene analysis in osteoporosis and familial osteoporosis. Osteoporos Int 15, 948–956 (2004). https://doi.org/10.1007/s00198-004-1654-x

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