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 Movement Cell Movement Cell Proliferation Cell Proliferation Down-Regulation Endometriosis Endometriosis Female Homeodomain Proteins Homeodomain Proteins Homeodomain Proteins Humans Kruppel-Like Factor 4

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