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

REPRODUCTIVE BIOMEDICINE ONLINE

卷:32

Article

Eggers, Julia C.¹  Martino, Valentina²  Reinbold, Rolland²  Schaefer, Sebastian D.¹  Kiesel, Ludwig¹  Starzinski-Powitz, Anna³  Schuering, Andreas N.¹  Kemper, Bjoern⁴  Greve, Burkhard⁵  Goette, Martin¹ 

[1] Munster Univ Hosp, Dept Gynecol & Obstet, D-48149 Munster, Germany

[2] CNR, Inst Biomed Technol, I-20090 Segrate, Italy

[3] Goethe Univ Frankfurt, Inst Anthropol & Human Genet Biologists, D-60054 Frankfurt, Germany

[4] Univ Munster, Biomed Technol Ctr, D-48149 Munster, Germany

[5] Univ Klinikum Munster, Klin Strahlentherapie Radioonkol, D-48149 Munster, Germany

关键词: EMT;    endometriosis;    microRNA;    miR-200b;    stem cells;    subfertility;   

DOI  :  10.1016/j.rbmo.2015.12.013

来源: Elsevier

【 摘 要 】

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. (C) 2016 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

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