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Reprod Biomed Online. 2016 Apr;32(4):434-45. doi: 10.1016/j.rbmo.2015.12.013. Epub 2016 Jan 15.

microRNA miR-200b affects proliferation, invasiveness and stemness of endometriotic cells by targeting ZEB1, ZEB2 and KLF4.

Author information

1
Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany.
2
Institute of Biomedical Technologies, National Research Council, 20090 Segrate-Milan, Italy.
3
Institute of Anthropology and Human Genetics for Biologists, Johann-Wolfgang-Goethe University of Frankfurt, Frankfurt, Germany.
4
Biomedical Technology Center, University of Muenster, 48149 Muenster, Germany.
5
Klinik für Strahlentherapie - Radioonkologie, Universitätsklinikum Münster, D-48149 Münster, Germany. Electronic address: greveb@uni-muenster.de.
6
Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany. Electronic address: mgotte@uni-muenster.de.

Abstract

Endometriosis is characterized by growth of endometrial tissue at ectopic locations. Down-regulation of microRNA miR-200b is observed in endometriosis and malignant disease, driving tumour cells towards an invasive state by enhancing epithelial-to-mesenchymal transition (EMT). miR-200b up-regulation may inhibit EMT and invasive growth in endometriosis. To study its functional impact on the immortalized endometriotic cell line 12Z, the stromal cell line ST-T1b, and primary endometriotic stroma cells, a transient transfection approach with microRNA precursors was employed. Expression of bioinformatically predicted targets of miR-200b was analysed by qPCR. The cellular phenotype was monitored by Matrigel invasion assays, digital-holographic video microscopy and flow cytometry. qPCR revealed significant down-regulation of ZEB1 (P < 0.05) and ZEB2 (P < 0.01) and an increase in E-cadherin (P < 0.01). miR-200b overexpression decreased invasiveness (P < 0.0001) and cell motility (P < 0.05). In contrast, cell proliferation (P < 0.0001) and the stemness-associated side population phenotype (P < 0.01) were enhanced following miR-200b transfection. These properties were possibly due to up-regulation of the pluripotency-associated transcription factor KLF4 (P < 0.05) and require attention when considering therapeutic strategies. In conclusion, up-regulation of miR-200b reverts EMT, emerging as a potential therapeutic approach to inhibit endometriotic cell motility and invasiveness.

KEYWORDS:

EMT; endometriosis; miR-200b; microRNA; stem cells; subfertility

PMID:
26854065
DOI:
10.1016/j.rbmo.2015.12.013
[Indexed for MEDLINE]

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