scholarly journals Loss of miR-542-3p enhances IGFBP-1 expression in decidualizing human endometrial stromal cells

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Hideno Tochigi ◽  
Takeshi Kajihara ◽  
Yosuke Mizuno ◽  
Yumi Mizuno ◽  
Shunsuke Tamaru ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Uterine Fibroids ◽  
Premenopausal Women ◽  
Hormonal Treatment ◽  
Adenosine Monophosphate ◽  
Luciferase Reporter ◽  
Marker Genes ◽  
Morphological Transformation ◽  
Endometrial Stromal Cells

Abstract Endometrial decidualization represents an essential step for the successful implantation of the embryo; however, the molecular mechanism behind this differentiation process remains unclear. This study aimed to identify novel microRNAs (miRNAs) involved in the regulation of decidual gene expression in human endometrial stromal cells (HESCs). An in vitro analysis of primary undifferentiated and decidualizing HESCs was conducted. HESCs were isolated from hysterectomy specimens from normally cycling premenopausal women with uterine fibroids, who were not on hormonal treatment at the time of surgery. Primary HESCs were expanded in culture and decidualized with 8-bromo-cyclic adenosine monophosphate and medroxyprogesterone acetate. Microarray analysis identified six miRNAs differentially expressed in response to decidualization of HESCs. All but one miRNA were downregulated upon decidualization, including miR-542-3p. We demonstrated that miR-542-3p overexpression inhibits the induction of major decidual marker genes, including IGFBP1, WNT4 and PRL. In addition, miR-542-3p overexpression inhibited the morphological transformation of HESCs in response to deciduogenic cues. A luciferase reporter assay confirmed that the 3′-untranslated region of IGFBP1 mRNA is targeted by miR-542-3p. The results suggest that miR-542-3p plays an important role in endometrial decidualization by regulating the expression of major decidual marker genes.

scholarly journals MiR-133b Improves Decidualization of Endometrial Stromal Cells by Targeting KLF12 in Recurrent Implantation Failure

2021 ◽  
Author(s):  
Fenglin Mei ◽  
Chengcai Kong ◽  
Yan Wang ◽  
Jing Zhuang ◽  
Pingping Xue ◽  
...  
Keyword(s):  
Western Blot ◽  
Stromal Cells ◽  
Luciferase Reporter ◽  
Implantation Failure ◽  
Reporter System ◽  
Recurrent Implantation Failure ◽  
Endometrial Stromal Cells ◽  
Protein Levels ◽  
Qrt Pcr

Abstract Purpose Impaired decidualization contributes to the infertility in recurrent implantation failure (RIF). Herein, we focused on the function and probable mechanisms of miR-133b in endometrial stromal cells decidualization.Methods miR-133b and KLF12 protein levels in midsecretory endometrial tissues derived from women with and without RIF were measured by qRT-PCR and Western blot. Primary human endometrial stromal cells (HESCs) were isolated and cultured for in vitro decidualization assays. Luciferase reporter, qRT-PCR and Western blot assays were used to measure the relationship between miR-133b and KLF12.Results miR-133b was significantly downregulated, whereas KLF12 was upregulated in endometrial tissues from RIF. miR-133b effectively promoted HESCs in vitro decidualization through the modulation of KLF12 expression and the activation of LIF/STAT3 pathway. Conversely, inhibition of miR-133b expression reversed these effects. In addition, the luciferase reporter system demonstrated that miR-133b directly inhibited the expression of KLF12 by interacting with 3’ untranslated region of KLF12.Conclusion Our data suggest that miR-133b promotes HESCs decidualization by targeting KLF12 and reverses the impaired decidualization in RIF.

scholarly journals Decidualization is Impaired in Endometrial Stromal Cells from Uterine Rudiments in Mayer-Rokitansky-Küster-Hauser Syndrome

2017 ◽  
Vol 41 (3) ◽  
pp. 1083-1097 ◽  
Author(s):  
Sara Y. Brucker ◽  
Simone Eisenbeis ◽  
Juliana König ◽  
Melanie Lamy ◽  
Madhuri S. Salker ◽  
...  
Keyword(s):  
Stromal Cells ◽  
University Hospital ◽  
Marker Genes ◽  
Proliferative Capacity ◽  
Receptor Function ◽  
Hormonal Stimulation ◽  
Endometrial Stromal Cells ◽  
Potential Sign

Background/Aims: Uterine rudiments from patients with Mayer-Rokitansky-Küster-Hauser syndrome (MRKHS) contain all tissues typically found in the uterus. Endometrium from the rudiments predominantly exhibits basalis-like features, and endometrial proliferative capacity in patients’ epithelium and stroma is significantly lower. Methods: This single-center, prospective study conducted at a major German university hospital compared in-vitro decidualization in cultured ESCs from MRKHS patients and hysterectomy controls. Primary ESC cultures were established from both sources. Hormone-induced prolactin and IGFBP-1 secretion served as a measure of their ability to undergo decidualization in response to hormonal stimulation. Expression levels of 8 key marker genes of decidualization were also determined. Results: At day 9, mean secretion of prolactin and IGFBP-1 was significantly reduced by 89.0% and 99.5%, respectively, in MRKHS ESCs vs. hysterectomy controls, both indicating impaired decidualization of MRKHS ESCs. Key decidual markers confirmed impaired decidualization in MRKHS patients. Conclusion: Our results indicate that the ESCs from MRKHS patients lack hormone responsiveness as a potential sign of dysfunctional hormone receptor function, which may also play a role in the onset of MRKHS. Further studies are needed to corroborate our findings, directly address receptor function, and elucidate the role of other potential determinants of uterine development and adult function.

scholarly journals Progesterone Receptor Transcriptome and Cistrome in Decidualized Human Endometrial Stromal Cells

Endocrinology ◽  
2015 ◽  
Vol 156 (6) ◽  
pp. 2239-2253 ◽  
Author(s):  
Erik C. Mazur ◽  
Yasmin M. Vasquez ◽  
Xilong Li ◽  
Ramakrishna Kommagani ◽  
Lichun Jiang ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Stromal Cells ◽  
Cellular Proliferation ◽  
Direct Interaction ◽  
Marker Genes ◽  
Regulation Of Transcription ◽  
Activator Protein 1 ◽  
Endometrial Stromal Cells ◽  
Complex Process

Abstract Decidualization is a complex process involving cellular proliferation and differentiation of the endometrial stroma that is required to establish and support pregnancy. Progesterone acting via its nuclear receptor, the progesterone receptor (PGR), is a critical regulator of decidualization and is known to interact with certain members of the activator protein-1 (AP-1) family in the regulation of transcription. In this study, we identified the cistrome and transcriptome of PGR and identified the AP-1 factors FOSL2 and JUN to be regulated by PGR and important in the decidualization process. Direct targets of PGR were identified by integrating gene expression data from RNA sequencing with the whole-genome binding profile of PGR determined by chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) in primary human endometrial stromal cells exposed to 17β-estradiol, medroxyprogesterone acetate, and cAMP to promote in vitro decidualization. Ablation of FOSL2 and JUN attenuates the induction of 2 decidual marker genes, IGFBP1 and PRL. ChIP-seq analysis of genomic binding revealed that FOSL2 is bound in proximity to 8586 distinct genes, including nearly 80% of genes bound by PGR. A comprehensive assessment of the PGR-dependent decidual transcriptome integrated with the genomic binding of PGR identified FOSL2 as a potentially important transcriptional coregulator of PGR via direct interaction with regulatory regions of genes actively regulated during decidualization.

scholarly journals Endometrial regeneration with endometrial epithelium: homologous orchestration with endometrial stroma as a feeder

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ryo Yokomizo ◽  
Yukiko Fujiki ◽  
Harue Kishigami ◽  
Hiroshi Kishi ◽  
Tohru Kiyono ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Therapeutic Option ◽  
Three Dimensional ◽  
Fertility Treatment ◽  
Feeder Cells ◽  
Endometrial Stromal Cells ◽  
Thin Endometrium ◽  
Endometrial Epithelial Cells

Abstract Background Thin endometrium adversely affects reproductive success rates with fertility treatment. Autologous transplantation of exogenously prepared endometrium can be a promising therapeutic option for thin endometrium; however, endometrial epithelial cells have limited expansion potential, which needs to be overcome in order to make regenerative medicine a therapeutic strategy for refractory thin endometrium. Here, we aimed to perform long-term culture of endometrial epithelial cells in vitro. Methods We prepared primary human endometrial epithelial cells and endometrial stromal cells and investigated whether endometrial stromal cells and human embryonic stem cell-derived feeder cells could support proliferation of endometrial epithelial cells. We also investigated whether three-dimensional culture can be achieved using thawed endometrial epithelial cells and endometrial stromal cells. Results Co-cultivation with the feeder cells dramatically increased the proliferation rate of the endometrial epithelial cells. We serially passaged the endometrial epithelial cells on mouse embryonic fibroblasts up to passage 6 for 4 months. Among the human-derived feeder cells, endometrial stromal cells exhibited the best feeder activity for proliferation of the endometrial epithelial cells. We continued to propagate the endometrial epithelial cells on endometrial stromal cells up to passage 5 for 81 days. Furthermore, endometrial epithelium and stroma, after the freeze-thaw procedure and sequential culture, were able to establish an endometrial three-dimensional model. Conclusions We herein established a model of in vitro cultured endometrium as a potential therapeutic option for refractory thin endometrium. The three-dimensional culture model with endometrial epithelial and stromal cell orchestration via cytokines, membrane-bound molecules, extracellular matrices, and gap junction will provide a new framework for exploring the mechanisms underlying the phenomenon of implantation. Additionally, modified embryo culture, so-called “in vitro implantation”, will be possible therapeutic approaches in fertility treatment.

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