Skip to main content
SpringerLink
Log in

Analysis of Clusterin and Clusterin Receptors in the Endometrium and Clusterin Levels in Cervical Mucus of Endometriosis

  • Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

Clusterin (CLU) is expressed in tissues and body fluids and is altered in some pathologies. In endometriosis, a noninvasive test is still lacking, thus, we analyzed CLU in mucus samples of patients. Additionally, we investigated localization of CLU and the putative CLU receptors (apolipoprotein E receptor 2 [ApoER2], megalin, very low-density lipoprotein receptor [VLDLR], and transforming growth factor β receptor type I and II [TβR1/TβR2]). In mucus samples, CLU levels are modestly, but not significantly, higher in cases with endometriosis compared to cases without endometriosis, however, CLU levels are significantly (P =.02) reduced in patients with endometriosis receiving contraception compared to cases with endometriosis without contraception. Analysis of CLU and CLU receptors showed CLU mainly in the uterine epithelial cells in the majority of glands, but also in endothelial cells. Similarly, ApoER2 and TβR1 could also be found preferentially in the endometrial glands. Whereas ApoER2 staining was strong in the vessels, TβR1 was modestly expressed in vessels and muscle cells. In contrast, staining of VLDLR and TbR2 was modest in the glands but stronger in vessels and muscle cells. Megalin staining was faint in the glands. A similar pattern for these proteins could be observed in adenomyosis. We demonstrate for the first time high concentrations of CLU in mucus samples and significantly reduced CLU levels in cases with endometriosis receiving contraception compared to cases with endometriosis without contraception. Furthermore, we identified uterine epithelial and endothelial cells as the main source of CLU and found different preferential CLU receptor complexes on glands, vessels, and smooth muscle cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Fritz, IB, Burdzy, K, Setchell, B, Blaschuk, O Ram rete testis fluid contains a protein (clusterin) which influences cell-cell-interactions in vitro. Biol Reprod. 1983;28(5):1173–1188.

    CAS  PubMed  Google Scholar 

  2. Shannan, B, Seifert, M, Leskov, K, et al. Challenge and promise: roles for clusterin in pathogenesis, progression and therapy of cancer. Cell Death Differ. 2006;13(1):12–19.

    CAS  PubMed  Google Scholar 

  3. Jin, G, Howe, PH Regulation of clusterin gene expression by transforming growth factor β. J Biol Chem. 1997;272(42): 26620–26626.

    CAS  PubMed  Google Scholar 

  4. Jin, G, Howe, PH Transforming growth factor β regulates clusterin gene expression via modulation of transcription factor c-Fos. Eur J Biochem. 1999;263(2):534–542.

    CAS  PubMed  Google Scholar 

  5. Bettuzzi, S, Davalli, P, Davoli, S, et al. Genetic inactivation of ApoJ/clusterin: effects on prostate tumourigenesis and metastatic spread. Oncogene. 2009;28(49):4344–4352.

    CAS  PubMed  Google Scholar 

  6. Chayka, O, Corvetta, D, Dews, M, et al. Clusterin, a haploinsuffi-cient tumor suppressor gene in neuroblastomas. J Natl Cancer Inst. 2009;101(9):663–677.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Bailey, RW, Aronow, B, Harmony, JA, Griswold, MD Heat shock-initiated apoptosis is accelerated and removal of damaged cells is delayed in the testis of clusterin/ApoJ knock-out mice. Biol Reprod. 2002;66(4):1042–1053.

    CAS  PubMed  Google Scholar 

  8. Trougakis, IP, So, A, Jansen, B, Gleave, ME, Gonos, ES Silencing expression of the clusterin/apolipoprotein j gene in human cancer cells using small interfering RNA induces spontaneous apoptosis, reduced growth ability, and cell sensitization to genotoxic and oxidative stress. Cancer Res. 2004;64(4):1834–1842.

    Google Scholar 

  9. Mishima, K, Inoue, H, Nishiyama, T, et al. Transplantation of side population cells restore the function of damaged exocrine glands through clusterin. Stem Cells. 2012;30(9):1925–1937.

    PubMed  Google Scholar 

  10. Thomas-Tikhonenko, A, Viard-Leveugle, I, Dews, M, et al. Myc-transformed epithelial cells down-regulate clusterin, which inhibits their growth in vitro and carcinogenesis in vivo. Cancer Res. 2004;64(9):3126–3136.

    CAS  PubMed  Google Scholar 

  11. Lenferink, AE, Cantin, C, Nantel, A, et al. Transcriptome profiling of a TGF-β-induced epithelial-to-mesenchymal transition reveals extracellular clusterin as a target for therapeutic antibodies. Onco-gene. 2010;29(6):831–844.

    CAS  Google Scholar 

  12. Nuutinen, T, Suuronen, T, Kauppinen, A, Saliminen, A Clusterin: a forgotten player in Alzheimer’s disease. Brain Res Rev. 2009; 61(2):89–104.

    CAS  PubMed  Google Scholar 

  13. McLaughlin, L, Zhu, G, Mistry, M, et al. Apolipoprotein J/clusterin limits the severity of murine autoimmune myocarditis. J Clin Invest. 2000;106(9):1105–1113.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Lee, KB, Jeon, JH, Choi, I, Kwon, OY, et al. Clusterin, a novel modulator of TGF-β signaling, is involved in Smad2/3 stability. Biochem Biophys Res Commun. 2008;366(4):905–909.

    CAS  PubMed  Google Scholar 

  15. Reddy, KB, Karode, MC, Harmony, AK, Howe, PH Interaction of transforming growth factor β receptors with apolipoprotein J/clus-terin. Biochemistry. 1996;35(1):309–314.

    CAS  PubMed  Google Scholar 

  16. Omwandho, CO, Konrad, L, Halis, G, Oehmke, F, Tinneberg, HR Role of TGF-betas in normal human endometrium and endome-triosis. Hum Reprod. 2010;25(1):101–109.

    CAS  PubMed  Google Scholar 

  17. Chegini, N, Zhao, Y, Williams, RS, Flanders, KC Human uterine tissue throughout the menstrual cycle expresses transforming growth factor-β1 (TGFβ1), TGFβ2, TGFβ3, and TGFβ type II receptor messenger ribonucleic acid and protein and contains [125I]TGFb1-binding sites. Endocrinology. 1994; 135(1):439–449.

    CAS  PubMed  Google Scholar 

  18. Zlokovic, BV, Martel, CL, Matsubara, E, et al. Glycoprotein 330/megalin: probable role in receptor-mediated transport of apolipoprotein J alone and in a complex with Alzheimer disease amyloid beta at the blood-brain and blood-cerebrospinal barriers. Proc Natl Acad Sci U S A 1996; 93(9):4229–4234.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Lundgren, S, Carling, T, Hjälm, G, et al. Tissue distribution of human gp330/megalin, a putative Ca2+-sensing protein. J Histo-chem Cytochem. 1997;45(3):383–392.

    CAS  Google Scholar 

  20. Nykjaer, A, Dragun, D, Walther, D, et al. An endocytic pathway essential for renal uptake and activation of the steroid 25-(OH) vitamin D3. Cell. 1999;96(4):507–515.

    CAS  PubMed  Google Scholar 

  21. Hammes, A, Andreassen, TK, Spoelgen, R, et al. Role of endocy-tosis in cellular uptake of sex steroids. Cell. 2005;122(5):751–762.

    CAS  PubMed  Google Scholar 

  22. Trommsdorff, M, Gotthardt, M, Hiesberger, T, et al. Reeler/ disabled-like disruption of neuronal migration in knockout mice lacking the VLDL receptor and ApoE receptor 2. Cell. 1999; 97(6):689–701.

    CAS  PubMed  Google Scholar 

  23. Leeb, C, Eresheim, C, Nimpf, J Clusterin is a ligand for apolipo-protein E receptor 2 (ApoER2) and very low density lipoprotein receptor (VLDLR) and signals via the Reelin-signaling pathway. J Biol Chem. 2014;289(7):4161–4172.

    CAS  PubMed  Google Scholar 

  24. Burk, RF, Hill, KE, Olson, GE, et al. Deletion of apolipoprotein E receptor-2 in mice lowers brain selenium and causes severe neu-rological dysfunction and death when a low-selenium diet is fed. J Neurosci. 2007;27(23):6207–6211.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Frykman, PK, Brown, MS, Yamamoto, T, Goldstein, JL, Herz, J Normal plasma lipoproteins and fertility in gene-targeted mice homozygous for a disruption in the gene encoding very low den-sity lipoprotein receptor. Proc Natl Acad Sci U S A 1995;92(18): 8453–8457.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Argraves, WS, Morales, CR Immunolocalization of cubilin, mega-lin, apolipoprotein, J, and apolipoprotein A-I in the uterus and oviduct. Mol Reprod Dev. 2004;69(4):419–427.

    CAS  PubMed  Google Scholar 

  27. Fuzio, P, Valletti, A, Napoli, A, et al. Regulation of the expression of CLU isoforms in endometrial proliferative diseases. Int J Oncol. 2013;42(6):1929–1944.

    CAS  PubMed  Google Scholar 

  28. Brown, TL, Moulton, BC, Baker, VV, Mira, J, Harmony, JA Expression of apolipoprotein J in the uterus is associated with tissue remodeling. Biol Reprod. 1995;52(5):1038–1049.

    CAS  PubMed  Google Scholar 

  29. Ferrero, S, Gillott, DJ, Remorgida, V, et al. Proteomic analysis of peritoneal fluid in fertile and infertile women with endometriosis. J Reprod Med. 2009;54(1):32–40.

    CAS  PubMed  Google Scholar 

  30. Hackethal, A, Luck, C, von Hobe, AK, et al. A structured ques-tionnaire improves preoperative assessment of endometriosis patients: a retrospective analysis and prospective trial. Arch Gyne-col Obstet. 2011;284(5):1179–1188.

    Google Scholar 

  31. Andersen, CL, Schepeler, T, Thorsen, K, et al. Clusterin expression in normal mucosa and colorectal cancer. Mol Cell Proteomics. 2007;6(6):1039–1048.

    CAS  PubMed  Google Scholar 

  32. Konrad, L, Scheiber, JA, Völck-Badouin, E, et al. Alternative spli-cing of TGF-betas and their high-affinity receptors TβRI, TβRII and TβRIII (Betaglycan) reveal new variants in human prostatic cells. BMC Genomics. 2007;8:318.

    PubMed  PubMed Central  Google Scholar 

  33. Varghese, F, Bukhari, AB, Malhotra, R, De, A IHC Profiler: an open source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue sam-ples. Plos One. 2014;9(5):e96801.

    PubMed  PubMed Central  Google Scholar 

  34. Fuhrich, DG, Lessey, BA, Savaris, RF Comparison of HSCORE assessment of endometrial β3 integrin subunit expression with digital HSCORE using computerized image analysis (ImageJ). Anal Quant Cytopathol Histopathol. 2013;35(4):210–216.

    Google Scholar 

  35. Fassbender, A, Burney, RO, Dorien, FO, et al. Update on biomar-kers for the detection of endometriosis. BioMed Res Int. 2015; 2015(3):130854.

    PubMed  PubMed Central  Google Scholar 

  36. de Bruijn, HW, van Beeck Calkoen-Carpay, T, Jager, S, Duk, JM, Aalders, JG, Fleuren, GJ The tumor marker CA 125 is a common constituent of normal cervical mucus. Am J Obstet Gynecol. 1986; 154(5):1088–1091.

    PubMed  Google Scholar 

  37. Rivera, R, Yacobson, I, Grimes, D The mechanism of action of hormonal contraceptives and intrauterine contraceptive devices. Am J Obstet Gynecol. 1999;181(5 pt 1):1263–1269.

    CAS  PubMed  Google Scholar 

  38. Endrikat, J, Parke, S, Trummer, D, Serrani, M, Duijkers, I, Klipping, C Pituitary, ovarian and additional contraceptive effects of an estradiol-based combined oral contraceptive: results of a rando-mized, open-label study. Contraception. 2013;87(2):227–234.

    CAS  PubMed  Google Scholar 

  39. Grandi, G, Mueller, M, Bersinger, NA, Cagnacci, A, Volpe, A, McKinnon, B Does dienogest influence the inflammatory response of endometriotic cells? A systematic review. Inflamm Res. 2016;65(3):183–192.

    CAS  PubMed  Google Scholar 

  40. Vitiello, D, Pinard, R, Taylor, HS Gene expression profiling reveals putative HOXA10 downstream. Reprod Sci. 2008;15(5): 529–535.

    CAS  PubMed  Google Scholar 

  41. Ulrich, V, Konaniah, ES, Herz, J, et al. Genetic variants of ApoE and ApoER2 differentially modulate endothelial function. Proc Natl Acad Sci U S A 2014;111(37):13493–13498.

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Andersen, OM, Yeung, CH, Vorum, H, et al. Essential role of the apolipoprotein E receptor-2 in sperm development. J Biol Chem. 2003;275(26):23989–23995.

    Google Scholar 

  43. Ramesh, S, Morrell, CN, Tarango, C, et al. Antiphospholipid anti-bodies promote leukocyte-endothelial cell adhesion and thrombo-sis in mice by antagonizing eNOS via β2GPI and apoER2. J Clin Invest. 2011;121(1):120–131.

    CAS  PubMed  Google Scholar 

  44. Yakovlev, S, Mikhailenko, I, Cao, C, Zhang, L, Strickland, DK, Medved, L Identification of VLDLR as a novel endothelial cell receptor for fibrin that modulates fibrin-dependent transendothe-lial migration of leukocytes. Blood. 2012;119(2):637–644.

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Li, Q, Agno, JE, Edson, MA, Nagaraja, AK, Nagashima, T, Matzuk, MM Transforming growth factor β receptor type 1 is essential for female reproductive tract integrity and function. PLOS Genet. 2011;7(10):e1002320.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Gao, Y, Bayless, KJ, Li, Q TGFBR1 is required for mouse myo-metrial development. Mol Endocrinol. 2014;28(3):380–394.

    PubMed  PubMed Central  Google Scholar 

  47. Ikushima, H, Miyazono, K TGF-β signal transduction spreading to a wider field: a broad variety of mechanisms for context-dependent effects of TGF-β. Cell Tissue Res. 2012; 347(1):37–49.

    CAS  PubMed  Google Scholar 

  48. Lee, S, Hong, SW, Min, BH, et al. Essential role of clusterin in pancreas regeneration. Dev Dyn. 2011;240(3):605–615.

    CAS  PubMed  Google Scholar 

  49. Falgarone, G, Chiocchia, G Chapter 8: Clusterin: a multifacet protein at the crossroad of inflammation and autoimmunity. Adv Cancer Res. 2009;104:139–170.

    CAS  PubMed  Google Scholar 

  50. Vercellini, P, Viganò, P, Somigliana, E, Fedele, L Endometriosis: pathogenesis and treatment. Nat Rev Endocrinol. 2014;10(5): 261–275.

    CAS  PubMed  Google Scholar 

  51. Trougakis, IP The molecular chaperone apolipoprotein J/clusterin as a sensor of oxidative stress: implications in therapeutic approaches–a mini-review. Gerontology. 2013;59(6):514–523.

    Google Scholar 

  52. Carvalho, LF, Samadder, AN, Agarwal, A, Fernandes, LF, Abrão, MS Oxidative stress biomarkers in patients with endometriosis: systematic review. Arch Gynecol Obstet. 2012;286(4):1033–1040.

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Gynecology and Obstetrics, Department of Obstetrics and Gynecology, Medical Faculty, Justus-Liebig-University, Feulgenstr. 12, D-35392, Giessen, Germany

    Lutz Konrad PhD, Frank Oehmke MD, Eniko Berkes MD, Jörg Engel MD & Hans-Rudolf Tinneberg MD

  2. Team Gynecologic Endoscopy, Private Clinic, Hamburg, Germany

    Andreas Hackethal MD

Authors
  1. Lutz Konrad PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andreas Hackethal MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Frank Oehmke MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Eniko Berkes MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jörg Engel MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hans-Rudolf Tinneberg MD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lutz Konrad PhD.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Konrad, L., Hackethal, A., Oehmke, F. et al. Analysis of Clusterin and Clusterin Receptors in the Endometrium and Clusterin Levels in Cervical Mucus of Endometriosis. Reprod. Sci. 23, 1371–1380 (2016). https://doi.org/10.1177/1933719116641756

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1177/1933719116641756

Keywords

Search

Navigation