title: Decidualization and syndecan-1 knock down sensitize endometrial stromal cells to apoptosis induced by embryonic stimuli.
abstract: Human embryo invasion and implantation into the inner wall of the maternal uterus, the endometrium, is the pivotal process for a successful pregnancy. Whereas disruption of the endometrial epithelial layer was already correlated with the programmed cell death, the role of apoptosis of the subjacent endometrial stromal cells during implantation is indistinct. The aim was to clarify whether apoptosis plays a role in the stromal invasion and to characterize if the apoptotic susceptibility of endometrial stromal cells to embryonic stimuli is influenced by decidualization and Syndecan-1. Therefore, the immortalized human endometrial stromal cell line St-T1 was used to first generate a new cell line with a stable Syndecan-1 knock down (KdS1), and second to further decidualize the cells with progesterone. As a replacement for the ethically inapplicable embryo all cells were treated with the embryonic factors and secretion products interleukin-1β, interferon-γ, tumor necrosis factor-α, transforming growth factor-β1 and anti-Fas antibody to mimic the embryo contact. Detection of apoptosis was verified via Caspase ELISAs, PARP cleavage and Annexin V staining. Apoptosis-related proteins were investigated via antibody arrays and underlying signaling pathways were analyzed by Western blot. Non-decidualized endometrial stromal cells showed a resistance towards apoptosis which was rescinded by decidualization and Syndecan-1 knock down independent of decidualization. This was correlated with an altered expression of several pro- and anti-apoptotic proteins and connected to a higher activation of pro-survival Akt in non-differentiated St-T1 as an upstream mediator of apoptotis-related proteins. This study provides insight into the largely elusive process of implantation, proposing an important role for stromal cell apoptosis to successfully establish a pregnancy. The impact of Syndecan-1 in attenuating the apoptotic signal is particularly interesting in the light of an already described influence on pregnancy disorders and therefore might provide a useful clinical tool in the future to prevent pregnancy complications provoked by inadequate implantation.
authors.affiliation: Department of Obstetrics/Gynecology and Reproductive Endocrinology and Infertility (UniKiD), Medical Center University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Sarah Jean
authors.initials: SJ
authors.lastname: Boeddeker
authors.affiliation: Department of Obstetrics/Gynecology and Reproductive Endocrinology and Infertility (UniKiD), Medical Center University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Dunja Maria
authors.initials: DM
authors.lastname: Baston-Buest
authors.affiliation: Department of Obstetrics and Gynecology, Medical Center University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Fehm
authors.affiliation: Department of Obstetrics/Gynecology and Reproductive Endocrinology and Infertility (UniKiD), Medical Center University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Kruessel
authors.affiliation: Department of Obstetrics/Gynecology and Reproductive Endocrinology and Infertility (UniKiD), Medical Center University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Hess
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002460:Cell Line; D016207:Cytokines / Q000633:toxicity*; D004717:Endometrium / Q000166:cytology; D005260:Female; D055785:Gene Knockdown Techniques; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011247:Pregnancy; D011374:Progesterone / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053668:Syndecan-1 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D019014:fas Receptor / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000906:Antibodies; D016207:Cytokines; C491901:FAS protein, human; D053668:Syndecan-1; D051996:Transcription Factor RelA; D019014:fas Receptor; D011374:Progesterone; D011065:Poly(ADP-ribose) Polymerases; D051057:Proto-Oncogene Proteins c-akt; D051550:I-kappa B Kinase; D048031:JNK Mitogen-Activated Protein Kinases; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9
references.citation: Semin Reprod Med. 2007 Nov;25(6):445-53
references.pmid: 17960529
references.citation: J Biol Chem. 2006 Aug 25;281(34):24365-74
references.pmid: 16807246
references.citation: Exp Cell Res. 2002 Apr 1;274(2):235-45
references.pmid: 11900484
references.citation: Placenta. 1998 Jan;19(1):21-6
references.pmid: 9481781
references.citation: Endocrinology. 1996 Jun;137(6):2225-31
references.pmid: 8641169
references.citation: Histol Histopathol. 2007 Mar;22(3):227-34
references.pmid: 17163397
references.citation: Mol Hum Reprod. 2014 Jun;20(6):567-78
references.pmid: 24478463
references.citation: J Endocrinol. 2003 Sep;178(3):357-72
references.pmid: 12967329
references.citation: Oncogene. 2008 Apr 7;27(16):2276-88
references.pmid: 18391970
references.citation: Am J Obstet Gynecol. 1990 Nov;163(5 Pt 1):1430-7
references.pmid: 2240083
references.citation: Acta Obstet Gynecol Scand. 2011 Oct;90(10):1157-63
references.pmid: 21501125
references.citation: Hum Reprod. 2001 Sep;16(9):1802-8
references.pmid: 11527879
references.citation: Reprod Biol. 2013 Jun;13(2):113-21
references.pmid: 23719115
references.citation: ScientificWorldJournal. 2003 Dec 11;3:1327-31
references.pmid: 14755113
references.citation: Baillieres Best Pract Res Clin Obstet Gynaecol. 2000 Oct;14(5):815-26
references.pmid: 11023802
references.citation: Contraception. 2005 Apr;71(4):249-62
references.pmid: 15792644
references.citation: J Mol Endocrinol. 2006 Jun;36(3):389-98
references.pmid: 16720711
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Mol Hum Reprod. 1998 Jan;4(1):87-91
references.pmid: 9510017
references.citation: Prenat Diagn. 2012 Feb;32(2):131-7
references.pmid: 22418956
references.citation: Virchows Arch. 2013 Sep;463(3):445-58
references.pmid: 23807541
references.citation: Biol Reprod. 2000 Aug;63(2):430-9
references.pmid: 10906047
references.citation: Biol Reprod. 2009 May;80(5):848-59
references.pmid: 19164174
references.citation: Biol Reprod. 1997 Oct;57(4):936-42
references.pmid: 9314600
references.citation: Annu Rev Biochem. 1999;68:729-77
references.pmid: 10872465
references.citation: Reprod Biol Endocrinol. 2003 May 26;1:44
references.pmid: 12801416
references.citation: Hum Reprod Update. 2004 Jan-Feb;10(1):29-38
references.pmid: 15005462
references.citation: Br J Cancer. 1972 Aug;26(4):239-57
references.pmid: 4561027
references.citation: J Immunol. 1999 May 15;162(10):6053-9
references.pmid: 10229846
references.citation: Endocrinology. 2007 Oct;148(10):5020-9
references.pmid: 17640988
references.citation: Fertil Steril. 2007 Mar;87(3):657-63
references.pmid: 17123519
references.citation: Clin Cancer Res. 2004 Aug 1;10(15):5178-86
references.pmid: 15297422
references.citation: Curr Biol. 2001 Oct 2;11(19):R795-805
references.pmid: 11591341
references.citation: Reprod Biol Endocrinol. 2009;7:76
references.pmid: 19619280
references.citation: Mol Hum Reprod. 2004 Jan;10(1):33-41
references.pmid: 14665704
references.citation: Genes Dev. 1999 Feb 1;13(3):239-52
references.pmid: 9990849
references.citation: Gynecol Oncol. 2009 Sep;114(3):509-15
references.pmid: 19539355
references.citation: Cell. 1998 Aug 21;94(4):491-501
references.pmid: 9727492
references.citation: Int J Oncol. 2002 Jan;20(1):39-43
references.pmid: 11743640
references.citation: Cell Death Differ. 2002 Oct;9(10):1031-42
references.pmid: 12232790
references.citation: Biol Reprod. 2007 Jan;76(1):102-17
references.pmid: 17021345
references.citation: Int J Cancer. 2007 Aug 15;121(4):741-50
references.pmid: 17455248
references.citation: Iran J Reprod Med. 2013 May;11(5):399-404
references.pmid: 24639772
references.citation: Mol Hum Reprod. 2011 Feb;17(2):127-34
references.pmid: 20926603
references.citation: J Immunol. 2012 Mar 15;188(6):2914-21
references.pmid: 22308310
references.citation: Clin Exp Obstet Gynecol. 2013;40(1):45-8
references.pmid: 23724505
references.citation: Endocrinology. 1994 Sep;135(3):1272-9
references.pmid: 8070373
references.citation: EMBO J. 1995 Nov 15;14(22):5579-88
references.pmid: 8521815
references.citation: J Biol Chem. 1998 Dec 11;273(50):33533-9
references.pmid: 9837934
references.citation: Am J Pathol. 2012 Feb;180(2):693-702
references.pmid: 22138583
references.citation: Cancer. 2003 Aug 1;98(3):474-83
references.pmid: 12879463
references.citation: J Cell Sci. 2007 Dec 1;120(Pt 23):4126-33
references.pmid: 18003704
references.citation: Reprod Biomed Online. 2007;14 Spec No 1:101-9
references.pmid: 20483405
references.citation: Fertil Steril. 1991 May;55(5):952-7
references.pmid: 1827077
references.citation: Reprod Biol Endocrinol. 2010;8:133
references.pmid: 21044331
references.citation: Hum Reprod. 1998 Aug;13(8):2206-11
references.pmid: 9756297
references.citation: Histochem J. 2000 Jun;32(6):373-9
references.pmid: 10943852
references.citation: J Clin Endocrinol Metab. 2000 Jun;85(6):2334-8
references.pmid: 10852473
references.citation: Oncogene. 1998 Dec 24;17(25):3247-59
references.pmid: 9916987
references.citation: Endocrinology. 2000 Sep;141(9):3510-3
references.pmid: 10965925
references.citation: Am J Obstet Gynecol. 1991 Oct;165(4 Pt 1):853-7
references.pmid: 1951543
references.citation: J Perinat Med. 2014 May;42(3):329-38
references.pmid: 24222257
references.citation: Endocrinology. 2006 Dec;147(12):5662-75
references.pmid: 16946011
references.citation: Blood. 2002 Feb 15;99(4):1405-10
references.pmid: 11830493