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Molecular characterization of soluble factors from human menstrual effluent that induce epithelial to mesenchymal transitions in mesothelial cells

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Abstract

We have studied menstrual effluent in order to identify soluble menstrual factors that induce epithelial to mesenchymal transitions (EMT) in mesothelial cells. A variety of molecules, such as nitric oxide and its reaction products, proteases (i.e. matrix metalloproteinases, plasmin) and proteins and/or peptides (i.e. growth factors: b-fibroblast growth factor, epidermal growth factor, hepatocyte growth factor, transforming growth factor-β; cytokines: interleukin 1β, tumour necrosis factor-α [TNF-α]) may be involved in this process. We have demonstrated that TNF-α is involved in EMT, whereas the other molecules are not. Biochemical analysis has shown that the inducing menstrual factors are heat-labile molecules, are uncharged at neutral pH, have a molecular weight between 50–70 kDa (or are bound in complexes of that size) and are eluted in the albumin fraction during gel filtration chromatography. Further analysis of this fraction by using proteomics and mass spectrometry has led to the identification of α-enolase and haemoglobin whose inhibition partially prevents EMT. When antibodies against TNF-α, α-enolase and haemoglobin are combined, EMT is almost completely inhibited. Thus, the candidates for soluble menstrual factors that induce mesothelial EMT are TNF-α, α-enolase and haemoglobin.

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Acknowledgements

The authors thank Dr. C.G.M. Heijnen (Department of Pharmacology and Toxicology, Maastricht University) for supplying the peroxynitrite, Mr. Barry Jütten for his skilful support in chromatography, Mrs. G.C. de Veen and Miss N. Kisters for their excellent technical support and Mr. Freek Bouwman for his contributions to the mass spectrophotometry analysis (Maastricht Proteomic Center, Maastricht University).

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Authors and Affiliations

  1. Research Institute Growth and Development (GROW), Academic Hospital and Maastricht University, Maastricht, The Netherlands

    Ayşe Y. Demir, Patrick G. Groothuis, Gerard A. J. Dunselman, Johannes L. H. Evers & Anton F. P. M. de Goeij

  2. Department of Obstetrics and Gynaecology, Academic Hospital and Maastricht University, Maastricht, The Netherlands

    Patrick G. Groothuis, Gerard A. J. Dunselman & Johannes L. H. Evers

  3. Department of Biochemistry, Academic Hospital and Maastricht University, Maastricht, The Netherlands

    Leon Schurgers

  4. Department of Pathology, Academic Hospital and Maastricht University, Maastricht, The Netherlands

    Anton F. P. M. de Goeij

  5. Department of Clinical Chemistry, University Medical Center Utrecht, G03.550, P.O. Box 85500, 3508 GA, Utrecht, The Netherlands

    Ayşe Y. Demir

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  1. Ayşe Y. Demir
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  2. Patrick G. Groothuis
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Correspondence to Ayşe Y. Demir.

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Demir, A.Y., Groothuis, P.G., Dunselman, G.A.J. et al. Molecular characterization of soluble factors from human menstrual effluent that induce epithelial to mesenchymal transitions in mesothelial cells. Cell Tissue Res 322, 299–311 (2005). https://doi.org/10.1007/s00441-005-0002-6

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  • DOI: https://doi.org/10.1007/s00441-005-0002-6

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