Cornelia Schneider, M.Sc., Nadja Kässens, M.D., Burkhard Greve, Ph.D., Hebatallah Hassan, M.Sc., Andreas Schüring, M.D., Anna Starzinski-Powitz, Ph.D., Ludwig Kiesel, M.D.,Ph.D., Daniela G. Seidler, Ph.D., Martin Götte, Ph.D.
Volume 99, Issue 3, Pages 871-881.e1, 1 March 2013
To study the function of Syndecan-1 and its potential regulator miR-10b in endometriosis.
Experimental laboratory study.
University medical center.
The human endometriotic cell line 12Z was transiently transfected with Syndecan-1 (SDC1) siRNA or miR-10b precursors and investigated for changes in cell behavior and gene expression. 12Z and primary eutopic endometrial stroma cells of two ASRM-III endometriosis patients were transfected with miR-10b precursors to investigate posttranscriptional regulation of SDC1.
Main Outcome Measure(s):
qPCR, Western blotting, flow cytometry, 3’UTR luciferase assays and zymography were employed to measure miR-10b-dependent targeting of SDC1, and SDC1-dependent expression changes of proteases and interleukin-6. Altered cell behavior was monitored by matrigel invasion assays, MTT cell viability assays and mitogen activated protein kinase (MAPK) activation blots.
SDC1 knockdown inhibited matrigel invasiveness by >60%, but did not affect cell viability. Interleukin-6 secretion, MMP-9 expression and MMP-2 activity were reduced, while PAI-1 protein expression was upregulated. miR-10b overexpression significantly downregulated SDC1, reduced matrigel invasiveness by 20%, and cell viability by 14%, and decreased MAPK activation in response to hepatocyte growth factor.
SDC1, a target of miR-10b, inhibits epithelial endometriotic cell invasiveness through downregulation of metalloproteinase activity and interleukin-6.
Read the full text at: http://www.fertstert.org/article/S0015-0282(12)02399-0/fulltext