MicroRNA miR 145 inhibits proliferation invasiveness and stem cell phenotype of an in vitro endometriosis model by targeting multiple cytoskeletal elements and pluripotency factors
MicroRNA miR-145 inhibits proliferation, invasiveness, and the stem cell phenotype of endometriotic cells by targeting multiple cytoskeletal elements and pluripotency factors, emerging as a novel target for therapeutic approaches.
Marlene Adammek, B.Sc., Burkhard Greve, Ph.D., Nadja Kässens, M.D., Cornelia Schneider, M.Sc., Kathrin Brüggemann, B.Sc., Andreas N. Schüring, M.D., Anna Starzinski-Powitz, Ph.D., Ludwig Kiesel, M.D.,Ph.D., Martin Götte, Ph.D.
Volume 99, Issue 5, Pages 1346-1355.e5, April 2013
To study the function of miR-145, known to be dysregulated in endometriosis, and to identify its target genes in an in vitro endometriosis model.
Experimental laboratory study.
University medical centers.
Primary endometrial stroma cells were derived from eutopic endometrium of three ASRMIII endometriosis patients and from ectopic lesions of four patients with deep infiltrating endometriosis.
The human endometriotic cell line 12Z and primary eutopic and ectopic endometrial stroma cells were transiently transfected with miR-145 precursors or antimiR-145 inhibitors and investigated for posttranscriptional regulation of predicted target genes and changes in cell behavior.
Main Outcome Measure(s):
Predicted target expression was measured by qRT-PCR and Western blotting, and altered cell behaviour was monitored by cell proliferation assays. 12Z cells were additionally investigated by matrigel invasion assays, cell cycle analysis, side population analysis and ALDH activity assays.
In all cells investigated, miR-145 overexpression inhibited cell proliferation, and induced downregulation of FASCIN-1, SOX2 and MSI2. In 12Z cells miR-145 upregulation increased matrigel invasiveness, and reduced side population and ALDH1 activity. Additional downregulated targets in 12Z cells included OCT4, KLF4, PODXL, JAM-A, and SERPINE1/PAI-1. ACTG2 and TAGLN were upregulated upon pre-miR-145 transfection. JAM-A, FASCIN-1 and PAI-I downregulation in 12Z cells were confirmed by Western blotting.
miR-145 inhibits endometriotic cell proliferation, invasiveness and stemness by targeting multiple pluripotency factors, cytoskeletal elements and protease inhibitors.