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

Julia C. Eggers, Eggers JC, Valentina Martino, Martino V, Rolland Reinbold, Reinbold R, Sebastian Daniel Schäfer, Sebastian D Schäfer, Ludwig Kiesel, Kiesel L, Anna Starzinski‐Powitz, Anna Starzinski-Powitz, Starzinski-Powitz A, Andreas N. Schüring, Björn Kemper, Burkhard Greve, Greve B, Martin Götte
Reproductive BioMedicine Online · 2016 · vol. 32(4) , pp. 434–445 · doi:10.1016/j.rbmo.2015.12.013 · PMID:26854065 · W2264241379
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This study investigated the effects of microRNA miR-200b on endometriotic cell proliferation, invasiveness, and stemness by examining its targeting of ZEB1, ZEB2, and KLF4.

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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.

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Condition tags

endometriosis

MeSH descriptors

Endometriosis Homeodomain Proteins Kruppel-Like Transcription Factors MicroRNAs Repressor Proteins Zinc Finger E-box-Binding Homeobox 1 Cell Line Cell Line Cell Movement Cell Movement Cell Movement Cell Proliferation Cell Proliferation Cell Proliferation Down-Regulation Down-Regulation Endometriosis Endometriosis Endometriosis Female

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