title: microRNA miR-200b affects proliferation, invasiveness and stemness of endometriotic cells by targeting ZEB1, ZEB2 and KLF4.
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.
authors: Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany.
authors: Julia C
authors: JC
authors: Eggers
authors: Institute of Biomedical Technologies, National Research Council, 20090 Segrate-Milan, Italy.
authors: Valentina
authors: V
authors: Martino
authors: Institute of Biomedical Technologies, National Research Council, 20090 Segrate-Milan, Italy.
authors: Rolland
authors: R
authors: Reinbold
authors: Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany.
authors: Sebastian D
authors: SD
authors: Schäfer
authors: Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany.
authors: Ludwig
authors: L
authors: Kiesel
authors: Institute of Anthropology and Human Genetics for Biologists, Johann-Wolfgang-Goethe University of Frankfurt, Frankfurt, Germany.
authors: Anna
authors: A
authors: Starzinski-Powitz
authors: Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany.
authors: Andreas N
authors: AN
authors: Schüring
authors: Biomedical Technology Center, University of Muenster, 48149 Muenster, Germany.
authors: Björn
authors: B
authors: Kemper
authors: Klinik für Strahlentherapie - Radioonkologie, Universitätsklinikum Münster, D-48149 Münster, Germany. Electronic address: greveb@uni-muenster.de.
authors: Burkhard
authors: B
authors: Greve
authors: Department of Gynecology and Obstetrics, Münster University Hospital, D-48149 Münster, Germany. Electronic address: mgotte@uni-muenster.de.
authors: Martin
authors: M
authors: Götte
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D015536:Down-Regulation; D004715:Endometriosis / Q000235:genetics* / Q000473:pathology; D005260:Female; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D035683:MicroRNAs / Q000502:physiology*; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D017154:Stromal Cells / Q000378:metabolism; D015854:Up-Regulation; D000074165:Zinc Finger E-box Binding Homeobox 2; D000071799:Zinc Finger E-box-Binding Homeobox 1 / Q000235:genetics* / Q000378:metabolism / Q000502:physiology
chemical_list: D018398:Homeodomain Proteins; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D012097:Repressor Proteins; C486307:ZEB1 protein, human; C492363:ZEB2 protein, human; D000074165:Zinc Finger E-box Binding Homeobox 2; D000071799:Zinc Finger E-box-Binding Homeobox 1
keywords: EMT; endometriosis; miR-200b; microRNA; stem cells; subfertility