TNFα and IL1β Stimulate Differential Gene Expression in Endometrial Stromal Cells
Author(s)
Abha J. Chalpe1,
Chad D. Law1,
Jennifer N. Dumdie1,
Keith A. Hansen2,
Kathleen M. Eyster1,2*
Affiliation(s)
1 Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, USA. 2 Department of Obstetrics and Gynecology, Sanford School of Medicine, The University of South Dakota, Sioux Falls, SD, USA.
1 Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, USA. 2 Department of Obstetrics and Gynecology, Sanford School of Medicine, The University of South Dakota, Sioux Falls, SD, USA.
ABSTRACT
The purpose of this study was to test the hypothesis that specific macrophage-secreted cytokines cause gene expression changes in endometrial stromal cells that reproduce the effects of macro-phages in the development of endometriosis. Telomerase-immortalized human endometrial stromal cells (T-HESC) were treated with tumor necrosis factor α (TNFα, 5 ng/ml) and interleukin 1β (IL1β, 1 ng/ml). Differential expression of 249 genes was identified by DNA microarray. Ontologies such as peptidases, cell adhesion, cell death/cell cycle, growth factors, cytoskeletal organization, defense/immune system, signal transduction, and transcriptional regulation which are related to the development of endometriosis were represented by these genes. The up-regulation of interleukin 8 (IL8), interleukin 6 (IL6), IL1β and matrix metallopro-teinase 3 (MMP3) in response to TNFα ± ILIβ in T-HESC cells was confirmed by real time RT-PCR. TNFα ± ILIβ did not affect the migration or invasion of T-HESC cells. This study reinforces our previous investigations on communication between cells of the immune system and endometrial stromal cells and their potential role in the development of endometriosis.
The purpose of this study was to test the hypothesis that specific macrophage-secreted cytokines cause gene expression changes in endometrial stromal cells that reproduce the effects of macro-phages in the development of endometriosis. Telomerase-immortalized human endometrial stromal cells (T-HESC) were treated with tumor necrosis factor α (TNFα, 5 ng/ml) and interleukin 1β (IL1β, 1 ng/ml). Differential expression of 249 genes was identified by DNA microarray. Ontologies such as peptidases, cell adhesion, cell death/cell cycle, growth factors, cytoskeletal organization, defense/immune system, signal transduction, and transcriptional regulation which are related to the development of endometriosis were represented by these genes. The up-regulation of interleukin 8 (IL8), interleukin 6 (IL6), IL1β and matrix metallopro-teinase 3 (MMP3) in response to TNFα ± ILIβ in T-HESC cells was confirmed by real time RT-PCR. TNFα ± ILIβ did not affect the migration or invasion of T-HESC cells. This study reinforces our previous investigations on communication between cells of the immune system and endometrial stromal cells and their potential role in the development of endometriosis.
KEYWORDS
Cytokines, Endometriosis, Endometrium, Endometrial Stromal Cells, Microarray, Gene Expression
Cytokines, Endometriosis, Endometrium, Endometrial Stromal Cells, Microarray, Gene Expression
Cite this paper
Chalpe, A. , Law, C. , Dumdie, J. , Hansen, K. and Eyster, K. (2015) TNFα and IL1β Stimulate Differential Gene Expression in Endometrial Stromal Cells. Advances in Biological Chemistry, 5, 126-141. doi: 10.4236/abc.2015.52010.
Chalpe, A. , Law, C. , Dumdie, J. , Hansen, K. and Eyster, K. (2015) TNFα and IL1β Stimulate Differential Gene Expression in Endometrial Stromal Cells. Advances in Biological Chemistry, 5, 126-141. doi: 10.4236/abc.2015.52010.
References
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http://dx.doi.org/10.1016/j.fertnstert.2007.01.056 [3] Siristatidis, C., Nissotakis, C., Chrelias, C., Iacovidou, H. and Salamalekis, E. (2006) Immunological Factors and Their Role in the Genesis and Development of Endometriosis. Journal of Obstetrics and Gynaecology Research, 32, 162-170.
http://dx.doi.org/10.1111/j.1447-0756.2006.00373.x [4] Banu, S.K., Lee, J., Starzinski-Powitz, A. and Arosh, J.A. (2008) Gene Expression Profiles and Functional Characterization of Human Immortalized Endometriotic Epithelial and Stromal Cells. Fertility and Sterility, 90, 972-987.
http://dx.doi.org/10.1016/j.fertnstert.2007.07.1358 [5] Eyster, K.M., Hansen, K.A., Winterton, E., Klinkova, O., Drappeau, D. and Mark-Kappeler, C.J. (2010) Reciprocal Communication between Endometrial Stromal Cells and Macrophages. Reproductive Sciences, 17, 809-822.
http://dx.doi.org/10.1177/1933719110371854 [6] Klinkova, O., Hansen, K.A., Winterton, E., Mark, C.J. and Eyster, K.M. (2010) Two-Way Communication between Endometrial Stromal Cells and Monocytes. Reproductive Sciences, 17, 125-136.
http://dx.doi.org/10.1177/1933719109348922 [7] Bersinger, N.A., Günthert, A.R., McKinnon, B., Johann, S. and Mueller, M.D. (2011) Dose-Response Effect of Interleukin (IL)-1β, Tumour Necrosis Factor (TNF)-α, and Interferon-γ on the in Vitro Production of Epithelial Neutrophil Activating Peptide-78 (ENA-78), IL-8, and IL-6 by Human Endometrial Stromal Cells. Archives of Gynecology and Obstetrics, 283, 1291-1296.
http://dx.doi.org/10.1007/s00404-010-1520-3 [8] Maybin, J.A., Critchley, H.O.D. and Jabbour, H.N. (2011) Inflammatory Pathways in Endometrial Disorders. Molecular and Cellular Endocrinology, 335, 42-51.
http://dx.doi.org/10.1016/j.mce.2010.08.006 [9] Giudice, L.C. and Kao, L.C. (2004) Endometriosis. Lancet, 364, 1789-1799.
http://dx.doi.org/10.1016/S0140-6736(04)17403-5 [10] Jones, C.J.P., Nardo, L.G., Littam P. and Fazleabas, A.T. (2009) Ultrastructure of Ectopic Peritoneal Lesions from Women with Endometriosis, Including Observations on the Contribution of Coelomic Mesothelium. Reproductive Sciences, 16, 43-55.
http://dx.doi.org/10.1177/1933719108324891 [11] Jovanovic, M. and Vicovac, L. (2009) Interleukin-6 Stimulates Cell Migration, Invasion and Integrin Expression in HTR-8/SVneo Cell Line. Placenta, 30, 320-328.
http://dx.doi.org/10.1016/j.placenta.2009.01.013 [12] Krikun, G., Nor, G., Alvero, A., Guller, S., Schatz, F., Sapi, E., Rahman, M., Caze, R., Qumsiyeh, M. and Lockwood, C.J. (2004) A Novel Immortalized Human Endometrial Stromal Cell Line with Normal Progestational Response. Endocrinology, 145, 2291-2296.
http://dx.doi.org/10.1210/en.2003-1606 [13] Mai, K.T., Yazdi, H.M., Perkins, D.G. and Parks, W. (1997) Pathogenetic Role of the Stromal Cells in Endometriosis and Adenomyosis. Histopathology, 30, 430-442.
http://dx.doi.org/10.1046/j.1365-2559.1997.4910725.x [14] Eyster, K.M. and Brannian, J.D. (2009) Gene Expression Profiling in the Aging Ovary. Methods in Molecular Biology, 590, 71-89.
http://dx.doi.org/10.1007/978-1-60327-378-7_5 [15] Hellemans, J., Mortier, G., De Paepe, A., Speleman, F. and Vandesompele, J. (2007) qBase Relative Quantification Framework and Software for Management and Automated Analysis of Real-Time Quantitative PCR Data. Genome Biology, 8, R19.
http://dx.doi.org/10.1186/gb-2007-8-2-r19 [16] Brazma, A., Hingamp, P., Quackenbush, J., Sherlock, G., Spellman, P., Stoeckert, C., Aach, J., Ansorge, W., Ball, C.A., Causton, H.C., et al. (2001) Minimum Information about a Microarray Experiment (MIAME)—Toward Standards for Microarray Data. Nature Genetics, 29, 365-371.
http://dx.doi.org/10.1038/ng1201-365 [17] Bondza, P.K., Maheux, R. and Akoum, A. (2009) Insights into Endometriosis-Associated Endometrial Dysfunctions: A Review. Frontiers in Bioscience, 1, 415-428. [18] Osteen, K.G., Keller, N.R., Feltus, F.A. and Melner, M.H. (1999) Paracrine Regulation of Matrix Metalloproteinase Expression in the Normal Human Endometrium. Gynecologic and Obstetric Investigation, 48, 2-13.
http://dx.doi.org/10.1159/000052863 [19] Braundmeier, A.G. and Nowak, R.A. (2006) Cytokines Regulate Matrix Metalloproteinases in Human Uterine Endometrial Fibroblast Cells through a Mechanism that Does Not Involve Increases in Extracellular Matrix Metalloproteinase Inducer. American Journal of Reproductive Immunology, 56, 201-214.
http://dx.doi.org/10.1111/j.1600-0897.2006.00418.x [20] Guay, S. and Akoum, A. (2007) Stable Inhibition of Inter-leukin 1 Receptor Type II in Ishikawa Cells Augments Secretion of Matrix Metalloproteinases: Possible Role in Endometriosis Pathophysiology. Reproduction, 134, 525-534.
http://dx.doi.org/10.1530/REP-06-0377 [21] Yang, J.H., Wu, M.Y., Chen, M.J., Chen, S.U., Yang, Y.S. and Ho, H.N. (2009) Increased Matrix Metalloproteinase-2 and Tissue Inhibitor of Met-alloproteinase-1 Secretion but Unaffected Invasiveness of Endometrial Stromal Cells in Adenomyosis. Fertility and Sterility, 91, 2193-2198.
http://dx.doi.org/10.1016/j.fertnstert.2008.05.090 [22] Milewski, L., Dziunycz, P., Barcz, E., Radomski, D., Roszkowski, P.I., Korczak-Kowalska, G., Kamiński, P. and Malejczyk, J. (2011) Increased Levels of Human Neutrophil Peptides 1, 2, and 3 in Peritoneal Fluid of Patients with Endometriosis: Association with Neutrophils, T Cells and IL-8. Journal of Reproductive Immunology, 91, 64-70.
http://dx.doi.org/10.1016/j.jri.2011.05.008 [23] Akoum, A., Lawson, C., McColl, S. and Villeneuve, M. (2001) Ectopic Endometrial Cells Express High Concentrations of Interleukin (IL)-8 in Vivo Regardless of the Menstrual Cycle Phase and Respond to Oestradiol by Up-Regulating IL-1-Induced IL-8 Expression in Vitro. Molecular Human Reproduction, 7, 859-866.
http://dx.doi.org/10.1093/molehr/7.9.859 [24] Lebovic, D.I., Bentzien, F., Chao, V.A., Garrett, E.N., Meng, Y.G. and Taylor, R.N. (2000) Induction of an Angiogenic Phenotype in Endometriotic Stromal Cell Cultures by Interleukin-1β. Molecular Human Reproduction, 6, 269-275.
http://dx.doi.org/10.1093/molehr/6.3.269 [25] Hou, Z., Zhou, J., Ma, X., Fan, L., Liao, L. and Liu, J. (2008) Role of Interleukin-1 Receptor Type II in the Pathogenesis of Endometriosis. Fertility and Sterility, 89, 42-51.
http://dx.doi.org/10.1016/j.fertnstert.2007.01.044 [26] Ponce, C., Torres, M., Galleguillos, C., Sovino, H., Boric, M.A., Fuentes, A. and Johnson, M.C. (2009) Nuclear Factor κB Pathway and Interleukin-6 Are Affected in Eutopic Endometrium of Women with Endometriosis. Reproduction, 137, 727-737.
http://dx.doi.org/10.1530/REP-08-0407 [27] Fassbender, A., Overbergh, L., Verdrengh, E., Kyama, C.M., Vodolazakaia, A., Bokor, A., Mueleman, C., Peeraer, K., Tomassetti, C., Waelkens, E., et al. (2011) How Can Macroscopically Normal Peritoneum Contribute to the Pathogenesis of Endometriosis? Fertility and Sterility, 96, 697-699.
http://dx.doi.org/10.1016/j.fertnstert.2011.06.034 [28] Shan, K., Wang, Y., Zhang, J.H., Guo, W., Wang, N. and Li, Y. (2005) The Function of the SNP in the MMP1 and MMP3 Promoter in Susceptibility to Endometriosis in China. Molecular Human Reproduction, 11, 423-427.
http://dx.doi.org/10.1093/molehr/gah177 [29] Sharpe-Timms, K.L., Nabli, H., Zimmer, R.L., Birt, J.A. and Davis, J.W. (2010) Inflammatory Cytokines Differentially Up-Regulate Human Endometrial Haptoglobin Production in Women with Endometriosis. Human Reproduction, 25, 1241-1250.
http://dx.doi.org/10.1093/humrep/deq032
[1] Kyama, C.M., Mihalyi, A., Simsa, P., Falconer, H., Fulop, V., Mwenda, J.M., Peeraer, K., Tomassetti, C., Meuleman, C. and D’Hooghe, T.M. (2009) Role of Cytokines in the Endometrial-Peritoneal Cross-Talk and Development of Endometriosis. Front Biosci (Elite Ed), 1, 444-454. [2] Eyster, K.M., Klinkova, O., Kennedy, V. and Hansen, K.A. (2007) Whole Genome Deoxyribonucleic Acid Microarray Analysis of Gene Expression in Ectopic versus Eutopic Endometrium. Fertility and Sterility, 88, 1505-1533.
http://dx.doi.org/10.1016/j.fertnstert.2007.01.056 [3] Siristatidis, C., Nissotakis, C., Chrelias, C., Iacovidou, H. and Salamalekis, E. (2006) Immunological Factors and Their Role in the Genesis and Development of Endometriosis. Journal of Obstetrics and Gynaecology Research, 32, 162-170.
http://dx.doi.org/10.1111/j.1447-0756.2006.00373.x [4] Banu, S.K., Lee, J., Starzinski-Powitz, A. and Arosh, J.A. (2008) Gene Expression Profiles and Functional Characterization of Human Immortalized Endometriotic Epithelial and Stromal Cells. Fertility and Sterility, 90, 972-987.
http://dx.doi.org/10.1016/j.fertnstert.2007.07.1358 [5] Eyster, K.M., Hansen, K.A., Winterton, E., Klinkova, O., Drappeau, D. and Mark-Kappeler, C.J. (2010) Reciprocal Communication between Endometrial Stromal Cells and Macrophages. Reproductive Sciences, 17, 809-822.
http://dx.doi.org/10.1177/1933719110371854 [6] Klinkova, O., Hansen, K.A., Winterton, E., Mark, C.J. and Eyster, K.M. (2010) Two-Way Communication between Endometrial Stromal Cells and Monocytes. Reproductive Sciences, 17, 125-136.
http://dx.doi.org/10.1177/1933719109348922 [7] Bersinger, N.A., Günthert, A.R., McKinnon, B., Johann, S. and Mueller, M.D. (2011) Dose-Response Effect of Interleukin (IL)-1β, Tumour Necrosis Factor (TNF)-α, and Interferon-γ on the in Vitro Production of Epithelial Neutrophil Activating Peptide-78 (ENA-78), IL-8, and IL-6 by Human Endometrial Stromal Cells. Archives of Gynecology and Obstetrics, 283, 1291-1296.
http://dx.doi.org/10.1007/s00404-010-1520-3 [8] Maybin, J.A., Critchley, H.O.D. and Jabbour, H.N. (2011) Inflammatory Pathways in Endometrial Disorders. Molecular and Cellular Endocrinology, 335, 42-51.
http://dx.doi.org/10.1016/j.mce.2010.08.006 [9] Giudice, L.C. and Kao, L.C. (2004) Endometriosis. Lancet, 364, 1789-1799.
http://dx.doi.org/10.1016/S0140-6736(04)17403-5 [10] Jones, C.J.P., Nardo, L.G., Littam P. and Fazleabas, A.T. (2009) Ultrastructure of Ectopic Peritoneal Lesions from Women with Endometriosis, Including Observations on the Contribution of Coelomic Mesothelium. Reproductive Sciences, 16, 43-55.
http://dx.doi.org/10.1177/1933719108324891 [11] Jovanovic, M. and Vicovac, L. (2009) Interleukin-6 Stimulates Cell Migration, Invasion and Integrin Expression in HTR-8/SVneo Cell Line. Placenta, 30, 320-328.
http://dx.doi.org/10.1016/j.placenta.2009.01.013 [12] Krikun, G., Nor, G., Alvero, A., Guller, S., Schatz, F., Sapi, E., Rahman, M., Caze, R., Qumsiyeh, M. and Lockwood, C.J. (2004) A Novel Immortalized Human Endometrial Stromal Cell Line with Normal Progestational Response. Endocrinology, 145, 2291-2296.
http://dx.doi.org/10.1210/en.2003-1606 [13] Mai, K.T., Yazdi, H.M., Perkins, D.G. and Parks, W. (1997) Pathogenetic Role of the Stromal Cells in Endometriosis and Adenomyosis. Histopathology, 30, 430-442.
http://dx.doi.org/10.1046/j.1365-2559.1997.4910725.x [14] Eyster, K.M. and Brannian, J.D. (2009) Gene Expression Profiling in the Aging Ovary. Methods in Molecular Biology, 590, 71-89.
http://dx.doi.org/10.1007/978-1-60327-378-7_5 [15] Hellemans, J., Mortier, G., De Paepe, A., Speleman, F. and Vandesompele, J. (2007) qBase Relative Quantification Framework and Software for Management and Automated Analysis of Real-Time Quantitative PCR Data. Genome Biology, 8, R19.
http://dx.doi.org/10.1186/gb-2007-8-2-r19 [16] Brazma, A., Hingamp, P., Quackenbush, J., Sherlock, G., Spellman, P., Stoeckert, C., Aach, J., Ansorge, W., Ball, C.A., Causton, H.C., et al. (2001) Minimum Information about a Microarray Experiment (MIAME)—Toward Standards for Microarray Data. Nature Genetics, 29, 365-371.
http://dx.doi.org/10.1038/ng1201-365 [17] Bondza, P.K., Maheux, R. and Akoum, A. (2009) Insights into Endometriosis-Associated Endometrial Dysfunctions: A Review. Frontiers in Bioscience, 1, 415-428. [18] Osteen, K.G., Keller, N.R., Feltus, F.A. and Melner, M.H. (1999) Paracrine Regulation of Matrix Metalloproteinase Expression in the Normal Human Endometrium. Gynecologic and Obstetric Investigation, 48, 2-13.
http://dx.doi.org/10.1159/000052863 [19] Braundmeier, A.G. and Nowak, R.A. (2006) Cytokines Regulate Matrix Metalloproteinases in Human Uterine Endometrial Fibroblast Cells through a Mechanism that Does Not Involve Increases in Extracellular Matrix Metalloproteinase Inducer. American Journal of Reproductive Immunology, 56, 201-214.
http://dx.doi.org/10.1111/j.1600-0897.2006.00418.x [20] Guay, S. and Akoum, A. (2007) Stable Inhibition of Inter-leukin 1 Receptor Type II in Ishikawa Cells Augments Secretion of Matrix Metalloproteinases: Possible Role in Endometriosis Pathophysiology. Reproduction, 134, 525-534.
http://dx.doi.org/10.1530/REP-06-0377 [21] Yang, J.H., Wu, M.Y., Chen, M.J., Chen, S.U., Yang, Y.S. and Ho, H.N. (2009) Increased Matrix Metalloproteinase-2 and Tissue Inhibitor of Met-alloproteinase-1 Secretion but Unaffected Invasiveness of Endometrial Stromal Cells in Adenomyosis. Fertility and Sterility, 91, 2193-2198.
http://dx.doi.org/10.1016/j.fertnstert.2008.05.090 [22] Milewski, L., Dziunycz, P., Barcz, E., Radomski, D., Roszkowski, P.I., Korczak-Kowalska, G., Kamiński, P. and Malejczyk, J. (2011) Increased Levels of Human Neutrophil Peptides 1, 2, and 3 in Peritoneal Fluid of Patients with Endometriosis: Association with Neutrophils, T Cells and IL-8. Journal of Reproductive Immunology, 91, 64-70.
http://dx.doi.org/10.1016/j.jri.2011.05.008 [23] Akoum, A., Lawson, C., McColl, S. and Villeneuve, M. (2001) Ectopic Endometrial Cells Express High Concentrations of Interleukin (IL)-8 in Vivo Regardless of the Menstrual Cycle Phase and Respond to Oestradiol by Up-Regulating IL-1-Induced IL-8 Expression in Vitro. Molecular Human Reproduction, 7, 859-866.
http://dx.doi.org/10.1093/molehr/7.9.859 [24] Lebovic, D.I., Bentzien, F., Chao, V.A., Garrett, E.N., Meng, Y.G. and Taylor, R.N. (2000) Induction of an Angiogenic Phenotype in Endometriotic Stromal Cell Cultures by Interleukin-1β. Molecular Human Reproduction, 6, 269-275.
http://dx.doi.org/10.1093/molehr/6.3.269 [25] Hou, Z., Zhou, J., Ma, X., Fan, L., Liao, L. and Liu, J. (2008) Role of Interleukin-1 Receptor Type II in the Pathogenesis of Endometriosis. Fertility and Sterility, 89, 42-51.
http://dx.doi.org/10.1016/j.fertnstert.2007.01.044 [26] Ponce, C., Torres, M., Galleguillos, C., Sovino, H., Boric, M.A., Fuentes, A. and Johnson, M.C. (2009) Nuclear Factor κB Pathway and Interleukin-6 Are Affected in Eutopic Endometrium of Women with Endometriosis. Reproduction, 137, 727-737.
http://dx.doi.org/10.1530/REP-08-0407 [27] Fassbender, A., Overbergh, L., Verdrengh, E., Kyama, C.M., Vodolazakaia, A., Bokor, A., Mueleman, C., Peeraer, K., Tomassetti, C., Waelkens, E., et al. (2011) How Can Macroscopically Normal Peritoneum Contribute to the Pathogenesis of Endometriosis? Fertility and Sterility, 96, 697-699.
http://dx.doi.org/10.1016/j.fertnstert.2011.06.034 [28] Shan, K., Wang, Y., Zhang, J.H., Guo, W., Wang, N. and Li, Y. (2005) The Function of the SNP in the MMP1 and MMP3 Promoter in Susceptibility to Endometriosis in China. Molecular Human Reproduction, 11, 423-427.
http://dx.doi.org/10.1093/molehr/gah177 [29] Sharpe-Timms, K.L., Nabli, H., Zimmer, R.L., Birt, J.A. and Davis, J.W. (2010) Inflammatory Cytokines Differentially Up-Regulate Human Endometrial Haptoglobin Production in Women with Endometriosis. Human Reproduction, 25, 1241-1250.
http://dx.doi.org/10.1093/humrep/deq032