scholarly journals Preimplantation human blastocysts release factors that differentially alter human endometrial epithelial cell adhesion and gene expression relative to IVF success

2013 ◽  
Vol 28 (5) ◽  
pp. 1161-1171 ◽  
Author(s):  
C. Cuman ◽  
E.M. Menkhorst ◽  
L.J. Rombauts ◽  
S. Holden ◽  
D. Webster ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Release Factors ◽  
Endometrial Epithelial Cell

scholarly journals Mouse double minute homologue 2 (MDM2) downregulation by miR-661 impairs human endometrial epithelial cell adhesive capacity

2018 ◽  
Vol 30 (3) ◽  
pp. 477 ◽  
Author(s):  
Amy Winship ◽  
Amanda Ton ◽  
Michelle Van Sinderen ◽  
Ellen Menkhorst ◽  
Katarzyna Rainczuk ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Endometrial Cells ◽  
Endometrial Epithelial Cell ◽  
Mouse Double Minute ◽  
Double Minute ◽  
Ishikawa Cells ◽  
Endometrial Epithelial Cells

Human blastocysts that fail to implant following IVF secrete elevated levels of miR-661, which is taken up by primary human endometrial epithelial cells (HEECs) and impairs their adhesive capability. MicroRNA miR-661 downregulates mouse double minute homologue 2 (MDM2) and MDM4 in other epithelial cell types to activate p53; however, this has not been examined in the endometrium. In this study MDM2 protein was detected in the luminal epithelium of the endometrium, the site of blastocyst attachment, during the mid secretory receptive phase of the menstrual cycle. The effects of miR-661 on gene expression in and adhesion of endometrial cells was also examined. MiR-661 overexpression consistently downregulated MDM2 but not MDM4 or p53 gene expression in the Ishikawa endometrial epithelial cell line and primary HEEC. Adhesion assays were performed on the real-time monitoring xCELLigence system and by co-culture using Ishikawa cells and HEECs with HTR8/SVneo trophoblast spheroids. Targeted siRNA-mediated knockdown of MDM2 in endometrial epithelial cells reduced Ishikawa cell adhesion (P < 0.001) and also reduced HTR8/SVneo trophoblast spheroid adhesion to Ishikawa cells (P < 0.05) and HEECs (P < 0.05). MDM2 overexpression using recombinant protein treatment resulted in enhanced HTR8/SVneo trophoblast spheroid adhesion to Ishikawa cells (P < 0.01) and HEECs (P < 0.05). This study highlights a potential new mechanism by which human blastocyst-secreted miR-661 reduces endometrial epithelial cell adhesion; via downregulation of MDM2. These findings suggest that MDM2 contributes to endometrial–blastocyst adhesion, implantation and infertility in women.

scholarly journals Human Blastocyst Secreted microRNA Regulate Endometrial Epithelial Cell Adhesion

EBioMedicine ◽  
2015 ◽  
Vol 2 (10) ◽  
pp. 1528-1535 ◽  
Author(s):  
Carly Cuman ◽  
Michelle Van Sinderen ◽  
Michael P. Gantier ◽  
Kate Rainczuk ◽  
Kelli Sorby ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Epithelial Cell ◽  
Endometrial Epithelial Cell ◽  
Human Blastocyst

scholarly journals Do GnRH analogues directly affect human endometrial epithelial cell gene expression?

2010 ◽  
Vol 16 (5) ◽  
pp. 347-360 ◽  
Author(s):  
X. Zhang ◽  
S. Bocca ◽  
A. Franchi ◽  
S. Anderson ◽  
M. Kaur ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cell ◽  
Gnrh Analogues ◽  
Endometrial Epithelial Cell ◽  
Cell Gene Expression ◽  
Cell Gene

scholarly journals Interleukin-11 and Leukemia Inhibitory Factor Regulate the Adhesion of Endometrial Epithelial Cells: Implications in Fertility Regulation

Endocrinology ◽  
2009 ◽  
Vol 150 (6) ◽  
pp. 2915-2923 ◽  
Author(s):  
M. Marwood ◽  
K. Visser ◽  
L. A. Salamonsen ◽  
E. Dimitriadis
Keyword(s):  
Cell Adhesion ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Leukemia Inhibitory Factor ◽  
Embryo Implantation ◽  
Neutralizing Antibody ◽  
Collagen Iv ◽  
Inhibitory Factor ◽  
Endometrial Epithelial Cell ◽  
Endometrial Epithelial Cells

Embryo implantation requires the closely harmonized processes of apposition, attachment, and adhesion of the conceptus to the maternal endometrial epithelium. IL-11 and leukemia inhibitory factor (LIF), two IL-6 family cytokines, are produced by the endometrium and are absolutely required for implantation in mice. We examined the effect of IL-11 and LIF on human endometrial epithelial cell adhesion. Both cytokines increased adhesion of primary human endometrial epithelial cells to fibronectin and collagen IV. IL-11 stimulated, whereas LIF had no effect on the adhesion of trophoblast to endometrial epithelial cells. Focused oligogene arrays were used to identify extracellular matrix and adhesion molecules mRNAs regulated by endometrial epithelial cells. We demonstrated by real-time RT-PCR and antibody arrays that both cytokines increased integrin-α2 mRNA and protein by endometrial epithelial cells. Signal transducers and activators of transcription (STAT)-3 inhibition reduced IL-11- and LIF-mediated epithelial cell adhesion to fibronectin, suggesting both cytokines regulated adhesion via phosphorylation of STAT3. Addition of either IL-11 neutralizing antibody and IL-11 or LIF and LIF antagonist to endometrial epithelial cells abolished cytokine induced phosphorylated STAT3. LIF but not IL-11 induced adhesion to collagen IV was reduced by an integrin-α2β1 neutralizing antibody. This study demonstrated that IL-11 and LIF regulated endometrial epithelial cell adhesion, suggesting that targeting IL-11 and LIF may be useful in regulating fertility by either enhancing or blocking implantation.

scholarly journals Solubility assessment of single-chain antibody fragment against epithelial cell adhesion molecule extracellular domain in four Escherichia coli strains

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Fatemeh Sadat Javadian ◽  
Majid Basafa ◽  
Aidin Behravan ◽  
Atieh Hashemi
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Extracellular Domain ◽  
Epithelial Cell Adhesion Molecule ◽  
Single Chain ◽  
E Coli ◽  
The Impact

Abstract Background Overexpression of the EpCAM (epithelial cell adhesion molecule) in malignancies makes it an attractive target for passive immunotherapy in a wide range of carcinomas. In comparison with full-length antibodies, due to the small size, the scFvs (single-chain variable fragments) are more suitable for recombinant expression in E. coli (Escherichia coli). However, the proteins expressed in large amounts in E. coli tend to form inclusion bodies that need to be refolded which may result in poor recovery of bioactive proteins. Various engineered strains were shown to be able to alleviate the insolubility problem. Here, we studied the impact of four E. coli strains on the soluble level of anti-EpEX-scFv (anti-EpCAM extracellular domain-scFv) protein. Results Although results showed that the amount of soluble anti-EpEX-scFv obtained in BL21TM (DE3) (114.22 ± 3.47 mg/L) was significantly higher to those produced in the same condition in E. coli RosettaTM (DE3) (71.39 ± 0.31 mg/L), and OrigamiTM T7 (58.99 ± 0.44 mg/L) strains, it was not significantly different from that produced by E. coli SHuffleTM T7 (108.87 ± 2.71 mg/L). Furthermore, the highest volumetric productivity of protein reached 318.29 ± 26.38 mg/L in BL21TM (DE3). Conclusions Although BL21TM (DE3) can be a suitable strain for high-level production of anti-EpEX-scFv protein, due to higher solubility yield (about 55%), E. coli SHuffleTM T7 seems to be better candidate for soluble production of scfv compared to BL21TM (DE3) (solubility yield of about 30%).

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