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Deregulation of LOXL1 and HTRA1 Gene Expression in Endometriosis

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Abstract

Endometriosis is a gynecologic disease characterized by the presence of endometrial tissue outside the uterine cavity. Although 15% of the female population in reproductive age is affected by endometriosis, its pathogenesis remains unclear. According to the most accepted pathogenesis hypothesis, endometrial fragments from the menstrual phase are transported through the uterine tubes to the peritoneal cavity, where they undergo implantation and growth, invading adjacent tissues. However, the establishment of the disease requires that endometrial cells present molecular characteristics favoring the onset and progression of ectopic implantation. In this investigation, we analyzed the differential gene expression profiles of peritoneal and ovarian endometriotic lesions compared to the endometrial tissue of nonaffected women using rapid subtraction hybridization (RaSH). In our study, this method was applied to samples of endometriotic lesions from affected women and to biopsies of endometrium of healthy women without endometriosis, where we could identify 126 deregulated genes. To evaluate the expression of genes found by RaSH method, we measured LOXL1, HTRA1, and SPARC genes by real-time polymerase chain reaction. Significant different expression was obtained for HTRA1 and LOXL1, upregulated in the ectopic endometrium, suggesting that these genes are involved in the physiopathology of endometriosis and may favor the viability of endometrial cells at ectopic sites.

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References

  1. Sharpe-Timms KL. Endometrial Anomalies in Women with Endometriosis. Ann N Y Acad Sci. 2001;943:131–147.

    Article  CAS  PubMed  Google Scholar 

  2. Yang WC, Chen HW, Au HK, et al. Serum and endometrial markers. Best Pract Res Clin Obstet Gynaecol. 2004;18(2):305–318.

    Article  PubMed  Google Scholar 

  3. Giudice LC, Kao LC. Endometriosis. Lancet. 2004;364(9447): 1789–1799.

    Article  PubMed  Google Scholar 

  4. Hassa H, Tanir HM, Uray M. Symptom distribution among infertile and fertile endometriosis cases with different stages and localizations. Eur J Obstet Gynecol Reprod Biol. 2005;119(1):82–86.

    Article  PubMed  Google Scholar 

  5. Redwine DB. Was Sampson wrong? Fertil Steril. 2002;78(4): 686–693.

    Article  PubMed  Google Scholar 

  6. Bischoff F, Simpson JL. Genetics of endometriosis: heritability and candidate genes. Best Pract Res Clin Obstet Gynaecol. 2004;18(2):219–232.

    Article  PubMed  Google Scholar 

  7. Arya P, Shaw R. Endometriosis: current thinking. Curr Obstet Gynaecol. 2005;15(3):191–198.

    Article  Google Scholar 

  8. Gaetje R, Kotzian S, Hermann G, Baumann R, Starzinski-Powitz A. Invasiveness of endometriotic cells in vitro. Lancet. 1995; 346(8988):1463–1464.

    Article  CAS  PubMed  Google Scholar 

  9. Viganò P, Somigliana E, Chiodo I, Abbiati A, Vercellini P. Molecular mechanisms and biological plausibility underlying the malignant transformation of endometriosis: a critical analysis. Hum Reprod Update. 2006;12(1):77–89.

    Article  PubMed  Google Scholar 

  10. Martelli M, Campana A, Bischof P. Secretion of matrix metalloproteinases by human endometrial cells in vitro. J Reprod Fertil. 1993;98(1):67–76.

    Article  CAS  PubMed  Google Scholar 

  11. Beliard A, Donnez J, Nisolle M, Foidart JM. Localization of laminin, fibronectin, E-cadherin, and integrins in endometrium and endometriosis. Fertil Steril. 1997;67(2):266–272.

    Article  CAS  PubMed  Google Scholar 

  12. Braun DP, Dmowski WP. Endometriosis: abnormal endometrium and dysfunctional immune response. Curr Opin Obstet Gynaecol. 1998;10(5):365–369.

    Article  CAS  Google Scholar 

  13. Healy DL, Rogers PA, Hii L, Wingfield M. Angiogenesis: a new theory for endometriosis. Hum Reprod Update. 1998;4(5): 736–740.

    Article  CAS  PubMed  Google Scholar 

  14. Zeitoun KM, Bulun SE. Aromatase: a key molecule in the pathophysiology of endometriosis. Fertil Steril. 1999;72(6):961–969.

    Article  CAS  PubMed  Google Scholar 

  15. Braun DP, Ding J, Shen J, Rana N, Fernandez BB, Dmowski WP. Relationship between apoptosis and the number of macrophages in eutopic endometrium from women with and without endometriosis. Fertil Steril. 2002;78(4):830–835.

    Article  PubMed  Google Scholar 

  16. Gogusev J, Bouquet de Joliniere J, Telvi L, et al. Detection of DNA copy number changes in human endometriosis by comparative genomic hybridization. Hum Genet. 1999;105(5):444–451.

    Article  CAS  PubMed  Google Scholar 

  17. Gogusev J, Bouquet de Joliniere J, Telvi L, et al. Genetic abnormalities detected by comparative genomic hybridization in a human endometriosis-derived cell line. Mol Hum Reprod. 2000;6(9): 821–827.

    Article  CAS  PubMed  Google Scholar 

  18. Kao LC, Germeyer A, Tulac S, et al. Expression profiling of endometrium from women with endometriosis reveals candidate genes for disease-based implantation failure and infertility. Endocrinology. 2003;144(7):2870–2881.

    Article  CAS  PubMed  Google Scholar 

  19. Guo SW, Wu Y, Strawnb E, et al. Genomic alterations in the endometrium may be a proximate cause for endometriosis. Eur J Obstet Gynaecol Reprod Biol. 2004;116(1):89–99.

    Article  CAS  Google Scholar 

  20. Flores I, Rivera E, Mousses S, Chen Y, Rozenblum E. Identification of molecular markers for endometriosis in blood lymphocytes by using deoxyribonucleic acid microarrays. Fertil Steril. 2006;85(6):1676–1683.

    Article  CAS  PubMed  Google Scholar 

  21. Honda H, Barrueto FF, Gogusev J, Im DD, Morin PJ. Serial analysis of gene expression reveals differential expression between endometriosis and normal endometrium. Possible roles for AXL and SHC1 in the pathogenesis of endometriosis. Reprod Biol Endocrinol. 2008;6:59.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. Meola J, Rosa e Silva JC, Dentillo DB, et al. Differentially expressed genes in eutopic and ectopic endometrium of women with endometriosis. Fertil Steril. 2010;93(6):1750–1773.

    Article  CAS  PubMed  Google Scholar 

  23. Eyster KM, Boles AL, Brannian JD, Hansen KA. DNA microarray analysis of gene expression markers of endometriosis. Fertil Steril. 2002;77(1):38–42.

    Article  PubMed  Google Scholar 

  24. Hu W, Tay SK, Zhao Y. Endometriosis-specific genes identified by real-time reverse transcription-polymerase chain reaction expression profiling of endometriosis versus autologous uterine endometrium. J Clin Endocrinol Metab. 2005; 91(1):228–238.

    Article  PubMed  CAS  Google Scholar 

  25. Kyama CM, Overbergh L, Debrock S, et al. Increased peritoneal and endometrial gene expression of biologically relevant cytokines and growth factors during the menstrual phase in women with endometriosis. Fertil Steril. 2006;85(6):1667–1675.

    Article  CAS  PubMed  Google Scholar 

  26. American Society for Reproductive Medicine. Revised American Society for Reproductive Medicine classification of endometriosis: 1996. Fertil Steril. 1997; 67(5):817–821.

    Article  Google Scholar 

  27. Jiang H, Kang DC, Alexandre D, Fisher PB. RaSH, a rapid subtraction hybridization approach for identifying and cloning differentially expressed genes. Proc Natl Acad Sci USA. 2000;97(23): 12684–12689.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25(4):402–408.

    CAS  PubMed  Google Scholar 

  29. Triola MF. Statdisk 9.1 for Elementary Statistics. 9th ed. Upper Saddle River, NJ: Pearson Education; 2005.

    Google Scholar 

  30. Lee SW, Tomasetto C, Sager R. Positive selection of candidate tumor-suppressor genes by subtractive hybridization. Proc Natl Acad Sci. 1991;88(7):2825–2829.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Sawada K, Kokeguchi S, Hongyo H, et al. Identification by Subtractive Hybridization of a Novel Insertion Sequence Specific for Virulent Strains of Porphyromonas gingivalis. Infect Immun. 1999;67(11):5621–5625.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Sha G, Wu D, Zhang L, et al. Differentially expressed genes in human endometrial endothelial cells derived from eutopic endometrium of patients with endometriosis compared with those from patients without endometriosis. Hum Reprod. 2007;22(12): 3159–3169.

    Article  CAS  PubMed  Google Scholar 

  33. Cho YJ. Identification of Up-Regulated Genes in Malignant Glioma with Subtraction Hybridization: Preliminary Screening Studies. Yonsei Med J. 2008;49(2):301–310.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Kawachi H, Suzuki K, Miyauchi N, Hashimoto T, Otaki Y, Shimizu F. Slit diaphragm dysfunction in proteinuric states: identification of novel therapeutic targets for nephrotic syndrome. Clin Exp Nephrol. 2009;13(4):275–280.

    Article  CAS  PubMed  Google Scholar 

  35. De Luca A, De Falco M, Severino A, et al. Distribution of the serine protease HtrA1 in normal human tissues. J Histochem Cytochem. 2003;51(10):1279–1284.

    Article  PubMed  Google Scholar 

  36. Bowden MA, Di Nezza-Cossens LA, Jobling T, Salamonsen LA, Nie G. Serine proteases HTRA1 and HTRA3 are down-regulated with increasing grades of human endometrial cancer. Gynecol Oncol. 2006;103(1):253–260.

    Article  CAS  PubMed  Google Scholar 

  37. Nie G, Hale K, Li Y, Manuelpillai U, Wallace EM, Salamonsen LA. Distinct Expression and Localization of Serine Protease HtrA1 in Human Endometrium and First-Trimester Placenta. Dev Dyn. 2006; 235(12):3448–3455.

    Article  CAS  PubMed  Google Scholar 

  38. Zumbrunn J, Trueb B. Primary structure of a putative serine protease specific for IGF-binding proteins. FEBS Lett. 1996; 398(2–3):187–192.

    Article  CAS  PubMed  Google Scholar 

  39. Clemmons DR. Role of insulin-like growth factor binding proteins in controlling IGF actions. Mol Cell Endocrinol. 1998;140(1–2):19–24.

    Article  CAS  PubMed  Google Scholar 

  40. Ferry RJ, Katz LE, Grimberg A, Cohen P, Weinzimer SA. Cellular actions of insulin-like growth factor binding proteins. Horm Metab Res. 1999;31(2–3):192–202.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Hou J, Clemmons DR, Smeekens S. Expression and characterization of a serine protease that preferentially cleaves insulin-like growth factor binding protein-5. J Cell Biochem. 2006;94(3):470–484.

    Article  CAS  Google Scholar 

  42. Gurgan T, Bukulmez O, Yarali H, Tanir M, Akyildiz S. Serum and peritoneal fluid levels of IGF I and II and insulinlike growth binding protein-3 in endometriosis. J Reprod Med. 1999;44(5): 450–454.

    CAS  PubMed  Google Scholar 

  43. Lembessis P, Milingos S, Michalas S, et al. Urokinase-type plasminogen activator and insulin-like growth factor-binding protein 3 mRNA expression in endometriotic lesions and eutopic endometrium: implications for the pathophysiology of endometriosis. Ann N Y Acad Sci. 2003;997:223–228.

    Article  CAS  PubMed  Google Scholar 

  44. Kagan HM, Li W. Lysyl oxidase: properties, specificity, and biological roles inside and outside of the cell. J Cell Biochem. 2003; 88(4):660–672.

    Article  CAS  PubMed  Google Scholar 

  45. Liu X, Zhao Y, Gao J, et al. Elastic fiber homeostasis requires lysyl oxidase-like 1 protein. Nat Genet. 2004;36(2):178–182.

    Article  CAS  PubMed  Google Scholar 

  46. Drewes PG, Yanagisawa H, Starcher B, et al. Pelvic organ prolapse in fibulin-5 knockout mice: pregnancy-induced changes in elastic fiber homeostasis in mouse vagina. Am J Pathol. 2007;170(2):578–589.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Talbi S, Hamilton AE, Vo KC, et al. Molecular phenotyping of human endometrium distinguishes menstrual cycle phases and underlying biological processes in normo-ovulatory women. Endocrinology. 2006;147(3):1097–1121.

    Article  CAS  PubMed  Google Scholar 

  48. Wu G, Guo Z, Chang X, et al. LOXL1 and LOXL4 are epigenetically silenced and can inhibit ras/extracellular signal-regulated kinase signaling pathway in human bladder cancer. Cancer Res. 2007;67(9):4123–4129.

    Article  CAS  PubMed  Google Scholar 

  49. Kirschmann DA, Seftor EA, Fong SF, et al. A molecular role for lysyl oxidase in breast cancer invasion. Cancer Res. 2002; 62(15): 4478–4483.

    CAS  PubMed  Google Scholar 

  50. Peinado H, Del Carmen Iglesias-de la Cruz M, Olmeda DA, et al. molecular role for lysyl oxidase-like 2 enzyme in Snail regulation and tumor progression. EMBO J. 2005;24(19):3446–3458.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Görögh T, Weise JB, Holtmeier C, et al. Selective upregulation and amplification of the lysyl oxidase like-4 (LOXL4) gene in head and neck squamous cell carcinoma. J Pathol. 2007;212(1): 74–82.

    Article  PubMed  CAS  Google Scholar 

  52. Görögh T, Holtmeier C, Weise JB, et al. Functional analysis of the 5’ flanking domain of the LOXL4 gene in head and neck squamous cell carcinoma cells. Int J Oncol. 2008;33(5):1091–1098.

    PubMed  Google Scholar 

  53. Flores I, Rivera E, Ruiz LA, Santiago OI, Vernon MW, Appleyard CB. Molecular profiling of experimental endometriosis identified gene expression patterns in common with human disease. Fertil Steril. 2007;87(5):1180–1199.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Konno R, Fujiwara H, Netsu S, et al. Gene expression profiling of the rat endometriosis model. Am J Reprod Immunol. 2007;58(4): 330–343.

    Article  CAS  PubMed  Google Scholar 

  55. Eyster KM, Klinkova O, Kennedy V, Hansen KA. Whole genome deoxyribonucleic acid microarray analysis of gene expression in ectopic versus eutopic endometrium. Fertil Steril. 2007;88(6): 1505–1533.

    Article  CAS  PubMed  Google Scholar 

  56. Bradshaw AD, Graves DC, Motamed K, Sage EH. SPARC-null mice exhibit increased adiposity without significant differences in overall body weight. Proc Natl Acad Sci USA. 2003;100(10): 6045–6050.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Poncelet AC, Schnaper HW. Regulation of human mesangial cell collagen expression by transforming growth factor-β1. Am J Physiol. 1998;275(3 pt 2):458–466.

    Google Scholar 

  58. Schiemann BJ, Neil JR, Schiemann WP. SPARC inhibits epithelial cell proliferation in part through stimulation of the transforming growth factor-β-signaling system. Mol Biol Cell. 2003;14(10):3977–3988.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Kato Y, Lewalle JM, Baba Y, et al. Induction of SPARC by VEGF in human vascular endothelial cells. Biochem Biophys Res Commun. 2001;287(2):422–426.

    Article  CAS  PubMed  Google Scholar 

  60. Chlenski A, Liu S, Guerrero LJ, et al. SPARC expression is associated with impaired tumor growth, inhibited angiogenesis and changes in the extracellular matrix. Int J Cancer. 2006;118(2): 310–316.

    Article  CAS  PubMed  Google Scholar 

  61. Campo McKnight DAC, Sosnoski DM, Koblinski JE, Gay CV. Roles of osteonectin in the migration of breast cancer cells into bone. J Cell Biochem. 2006; 97(2):288–302.

    Article  PubMed  CAS  Google Scholar 

  62. Smit DJ, Gardiner BB, Sturm RA. Osteonectin downregulates E-cadherin, induces osteopontin and focal adhesion kinase activity stimulating an invasive melanoma phenotype. Int J Cancer. 2007;121(12):2653–2660.

    Article  CAS  PubMed  Google Scholar 

  63. Goodsaid FM, Smith RJ, Rosenblum IY. Quantitative PCR deconstruction of discrepancies between results reported by different hybridization platforms. Environ Health Perspect. 2004; 112(4):456–460.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Cheng YH, Imir A, Fenkci V, Yilmaz MB, Bulun SE. Stromal cells of endometriosis fail to produce paracrine factors that induce epithelial 17beta-hydroxysteroid dehydrogenase type 2 gene and its transcriptional regulator Sp1: a mechanism for defective estradiol metabolism. Am J Obstet Gynecol. 2007;196(4):391.e1–391.e8.

    Article  CAS  Google Scholar 

  65. Mulayim N, Savlu A, Guzeloglu-Kayisli O, Kayisli UA, Arici A. Regulation of endometrial stromal cell matrix metalloproteinase activity and invasiveness by interleukin-8. Fertil Steril. 2004;81(suppl 1):904–911.

    Article  CAS  PubMed  Google Scholar 

  66. Malik S, Day K, Perrault I, Charnock-Jones DS, Smith SK. Menstrual effluent in endometriosis shows no difference in volume, VEGF-A, MMP2 and MMP9 or sFLT. Reprod Biomed Online. 2006;12(2):174–181.

    Article  CAS  PubMed  Google Scholar 

  67. Salata IM, Stojanovic N, Cajdler-Łuba A, Lewandowski KC, Lewiński A. Gelatinase A (MM-2), gelatinase B (MMP-9) and their inhibitors (TIMP 1, TIMP-2) in serum of women with endometriosis: significant correlation between MMP-2, MMP-9 and their inhibitors without difference in levels of matrix metalloproteinases and tissue inhibitors of metalloproteinases in relation to the severity of endometriosis. Gynecol Endocrinol. 2008;24(6):326–330.

    Article  CAS  PubMed  Google Scholar 

  68. Wang Y, Yu J, Luo X, et al. Abnormal regulation of chemokine TECK and its receptor CCR9 in the endometriotic milieu is involved in pathogenesis of endometriosis by way of enhancing invasiveness of endometrial stromal cells. Cell Mol. Immunol. 2010;7(1):51–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Depalo R, Cavallini A, Lorusso F, et al. Apoptosis in normal ovaries of women with and without endometriosis. Reprod Biomed Online. 2009;19(6):808–815.

    Article  CAS  PubMed  Google Scholar 

  70. Parra FC, Amado RC, Lambertucci JR, Rocha J, Antunes CM, Pena SD. Color and genomic ancestry in Brazilians. Proc Natl Acad Sci U S A. 2003;100(1):177–182.

    Article  CAS  PubMed  Google Scholar 

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Dentillo, D.B., Meola, J., Rosa e Silva, J.C. et al. Deregulation of LOXL1 and HTRA1 Gene Expression in Endometriosis. Reprod. Sci. 17, 1016–1023 (2010). https://doi.org/10.1177/1933719110377662

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