γ‐Secretase inhibition affects viability, apoptosis, and the stem cell phenotype of endometriotic cells

Williams, Laura Ramirez; Brüggemann, Kathrin; Hubert, Marina; Achmad, Nurjannah; Kiesel, L.; Schäfer, Sebastian D.; Greve, Burkhard; Götte, Martin

doi:10.1111/aogs.13707

Cited by 14 publications

(15 citation statements)

References 35 publications

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“…Based on previous findings in endometriosis linking MSI proteins to LIFR expression [30] we aimed to investigate this interplay in breast cancer. Similar to endometriosis, the LIFR was downregulated in breast cancer.…”

Section: Msi Protein Knockdown Results In a Higher Cell Invasiveness mentioning

confidence: 99%

Section: Msi-downregulated Cells Exhibit Increased Cell Motility and mentioning

confidence: 95%

“…Based on a previous study employing Notch inhibitors in endometriosis [30], we could show that the leukemia inhibitory factor receptor (LIFR) expression, a key inhibitor of cell invasion and motility, was downregulated in MSI knockdown cells both in qPCR (by more than 60%, p < 0.001, In colon cancer, MSI-1 expression and epithelial growth factor pathway activity were correlated [28], the latter being another key regulator of radioresistance that is specifically targeted in radiation-centered clinical trials [29]. As evidenced by our Western blot results, the epithelial growth factor receptor (EGFR) is downregulated following MSI knockdown (p < 0.05, Figure 4C, representative blot in Figure 4D).…”

Section: Msi-downregulated Cells Exhibit Increased Cell Motility and mentioning

confidence: 99%

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Knockdown of Musashi RNA Binding Proteins Decreases Radioresistance but Enhances Cell Motility and Invasion in Triple-Negative Breast Cancer

Troschel

Minte

Ismail

et al. 2020

IJMS

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The therapeutic potential of Musashi (MSI) RNA-binding proteins, important stemness-associated gene expression regulators, remains insufficiently understood in breast cancer. This study identifies the interplay between MSI protein expression, stem cell characteristics, radioresistance, cell invasiveness and migration. MSI-1, MSI-2 and Notch pathway elements were investigated via quantitative polymerase chain reaction (qPCR) in 19 triple-negative breast cancer samples. Measurements were repeated in MDA-MB-231 cells after MSI-1 and -2 siRNA-mediated double knockdown, with further experiments performed after MSI silencing. Flow cytometry helped quantify expression of CD44 and leukemia inhibitory factor receptor (LIFR), changes in apoptosis and cell cycle progression. Proliferation and irradiation-induced effects were assessed using colony formation assays. Radiation-related proteins were investigated via Western blots. Finally, cell invasion assays and digital holographic microscopy for cell migration were performed. MSI proteins showed strong correlations with Notch pathway elements. MSI knockdown resulted in reduction of stem cell marker expression, cell cycle progression and proliferation, while increasing apoptosis. Cells were radiosensitized as radioresistance-conferring proteins were downregulated. However, MSI-silencing-mediated LIFR downregulation resulted in enhanced cell invasion and migration. We conclude that, while MSI knockdown results in several therapeutically desirable consequences, enhanced invasion and migration need to be counteracted before knockdown advantages can be fully exploited.

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Section: Msi Protein Knockdown Results In a Higher Cell Invasiveness mentioning

confidence: 99%

Section: Msi-downregulated Cells Exhibit Increased Cell Motility and mentioning

confidence: 95%

Section: Msi-downregulated Cells Exhibit Increased Cell Motility and mentioning

confidence: 99%

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Knockdown of Musashi RNA Binding Proteins Decreases Radioresistance but Enhances Cell Motility and Invasion in Triple-Negative Breast Cancer

Troschel

Minte

Ismail

et al. 2020

IJMS

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“…ESCs were prepared from ectopic lesions and eutopic endometrium and characterized as previously described [28]. Primary endometriotic stromal cells were prepared from biopsies of women with endometriosis who underwent surgical treatment at the Department of Gynecology and Obstetrics of Münster University Hospital between 2009 and 2010 as well as from October 2012 and March 2014 as described previously [29]. The modified American Society for Reproductive Medicine classification was used to assess endometriosis (ASRM, 1997).…”

Section: Cell Culturementioning

confidence: 99%

miR-142-3p Reduces the Size, Migration, and Contractility of Endometrial and Endometriotic Stromal Cells by Targeting Integrin- and Rho GTPase-Related Pathways That Regulate Cytoskeletal Function

et al. 2020

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Downregulated microRNA-142-3p signaling contributes to the pathogenesis of endometriosis, an invasive disease where the lining of the uterus grows at ectopic locations, by yet incompletely understood mechanisms. Using bioinformatics and in vitro assays, this study identifies cytoskeletal regulation and integrin signaling as two relevant categories of miR-142-3p targets. qPCR revealed that miR-142-3p upregulation in St-T1b cells downregulates Rho-associated protein kinase 2 (ROCK2), cofilin 2 (CFL2), Ras-related C3 botulinum toxin substrate 1 (RAC1), neural Wiskott-Aldrich syndrome protein (WASL), and integrin α-V (ITGAV). qPCR and Western-blotting showed miR-142-3p effect on WASL and ITGAV was significant also in primary endometriotic stroma cells. Luciferase reporter assays in ST-T1b cells then confirmed direct regulation of ITGAV and WASL. On the functional side, miR-142-3p upregulation significantly reduced ST-T1b cell size, the size of vinculin plaques, migration through fibronectin-coated transwell filters, and the ability of ST-T1b and primary endometriotic stroma cells to contract collagen I gels. These results suggest that miR-142-3p has a strong mechanoregulatory effect on endometrial stroma cells and its external administration reduces the invasive endometrial phenotype. Within the limits of an in vitro investigation, our study provides new mechanistic insights into the pathogenesis of endometriosis and provides a perspective for the development of miR-142-3p based drugs for inhibiting invasive growth of endometriotic cells.

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“…Cells were routinely split twice a week. ESCs were prepared from ectopic lesions and characterized as previously described 58 . Primary endometriotic stroma cells were prepared from a biopsiy of a woman with endometriosis who underwent surgical treatment at the Department of Gynecology and Obstetrics of Münster University Hospital in 2013.…”

Section: Cell Culturementioning

confidence: 99%

The scar constituent Collagen I triggers coordinated collective migration and invasion in a 3D spheroid model of early endometriotic lesions

Stejskalová

Fincke

Nowak

et al. 2020

Preprint

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Endometriosis is a painful gynaecological condition characterized by ectopic growth of endometrial cells outside of the uterus. Little is known about the mechanisms by which endometrial fragments invade tissues. This is partially due to a lack of suitable experimental models. In this study, we show that a spheroid 3D model, but not single cells mimic the collective endometrial fragment-like invasion through the extracellular matrix. This model reveals that collagen I, the main constituent of surgical scars, significantly increases the rate of lesion formation by healthy endometrial stromal cells in vitro compared to the basement membrane-like matrix Matrigel. Stromal cell invasion of collagen I requires MMPs, whereas collective migration of endometriotic epithelial 12Z cells involves Rac-signalling. We show that inhibiting ROCK signalling responsible for actomyosin contraction increases the lesion-size. Moreover, endometriotic epithelial 12Z cells, but not eutopic stromal cells St-T1b migrate onMatrigel. The rate of this migration is decreased by the microRNA miR-200b and increased by miR-145. Our 3D model offers a facile approach to dissect how endometrial fragments invade tissues and is an important step toward developing new personalized therapeutics for endometriosis. Moreover, our model is a suitable tool to screen small molecule drugs and microRNA-based therapeutics.

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γ‐Secretase inhibition affects viability, apoptosis, and the stem cell phenotype of endometriotic cells

Cited by 14 publications

References 35 publications

Knockdown of Musashi RNA Binding Proteins Decreases Radioresistance but Enhances Cell Motility and Invasion in Triple-Negative Breast Cancer

Knockdown of Musashi RNA Binding Proteins Decreases Radioresistance but Enhances Cell Motility and Invasion in Triple-Negative Breast Cancer

miR-142-3p Reduces the Size, Migration, and Contractility of Endometrial and Endometriotic Stromal Cells by Targeting Integrin- and Rho GTPase-Related Pathways That Regulate Cytoskeletal Function

The scar constituent Collagen I triggers coordinated collective migration and invasion in a 3D spheroid model of early endometriotic lesions

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