Targeting of syndecan 1 by micro ribonucleic acid miR 10b modulates invasiveness of endometriotic cells via dysregulation of the proteolytic millieu and interleukin 6 secretion

Capsule:
Syndecan-1 inhibits invasiveness of epithelial endometriotic cells by modulating matrix metalloproteinases MMP-2 and MMP-9, plasminogen activator inhibitor-1, and interleukin-6 and is regulated by the microRNA miR-10b, both possible novel targets for therapeutic approaches.

Authors:
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

Abstract:

Objective:
To study the function of Syndecan-1 and its potential regulator miR-10b in endometriosis.

Design:
Experimental laboratory study.

Setting:
University medical center.

Patient(s):
Not applicable.

Intervention(s):
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.

Result(s):
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.

Conclusion(s):
SDC1, a target of miR-10b, inhibits epithelial endometriotic cell invasiveness through downregulation of metalloproteinase activity and interleukin-6.

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