Oncotarget

Research Papers:

Silencing of Profilin-1 suppresses cell adhesion and tumor growth via predicted alterations in integrin and Ca2+ signaling in T24M-based bladder cancer models

Maria Frantzi, Zoi Klimou, Manousos Makridakis, Jerome Zoidakis, Agnieszka Latosinska, Daniel M. Borràs, Bart Janssen, Ioanna Giannopoulou, Vasiliki Lygirou, Andreas C. Lazaris, Nicholas P. Anagnou, Harald Mischak _, Maria G. Roubelakis and Antonia Vlahou

PDF  |  HTML  |  Supplementary Files  |  How to cite  |  Order a Reprint

Oncotarget. 2016; 7:70750-70768. https://doi.org/10.18632/oncotarget.12218

Metrics: HTML 1201 views  |   ?  


Abstract

Maria Frantzi1,2, Zoi Klimou3,4, Manousos Makridakis1, Jerome Zoidakis1, Agnieszka Latosinska1, Daniel M. Borràs5, Bart Janssen5, Ioanna Giannopoulou6, Vasiliki Lygirou1, Andreas C. Lazaris6, Nicholas P. Anagnou3,4, Harald Mischak2, Maria G. Roubelakis3,4, Antonia Vlahou1

1Proteomics Laboratory, Biotechnology Division, Biomedical Research Foundation of the Academy of Athens, Athens, Greece

2Research and Development Department, Mosaiques Diagnostics GmbH, Hannover, Germany

3Laboratory of Biology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece

4Cell and Gene Therapy Laboratory, Biomedical Research Foundation of The Academy of Athens, Athens, Greece

5Research and Development Department, GenomeScan B.V., Leiden, The Netherlands

6First Department of Pathology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece

Correspondence to:

Maria G. Roubelakis, email: roubel@med.uoa.gr

Keywords: Profilin-1, bladder cancer, xenograft, gene silencing, metastasis

Received: March 22, 2016    Accepted: September 13, 2016    Published: September 23, 2016

ABSTRACT

Bladder cancer (BC) is the second most common malignancy of the genitourinary system, characterized by the highest recurrence rate of all cancers. Treatment options are limited; thus a thorough understanding of the underlying molecular mechanisms is needed to guide the discovery of novel therapeutic targets. Profilins are actin binding proteins with attributed pleiotropic functions to cytoskeletal remodeling, cell adhesion, motility, even transcriptional regulation, not fully characterized yet. Earlier studies from our laboratory revealed that decreased tissue levels of Profilin-1 (PFN1) are correlated with BC progression to muscle invasive disease. Herein, we describe a comprehensive analysis of PFN1 silencing via shRNA, in vitro (by employing T24M cells) and in vivo [(with T24M xenografts in non-obese diabetic severe combined immunodeficient mice (NOD/SCID) mice]. A combination of phenotypic and molecular assays, including migration, proliferation, adhesion assays, flow cytometry and total mRNA sequencing, as well as immunohistochemistry for investigation of selected findings in human specimens were applied. A decrease in BC cell adhesion and tumor growth in vivo following PFN downregulation are observed, likely associated with the concomitant downregulation of Fibronectin receptor, Endothelin-1, and Actin polymerization. A decrease in the levels of multiple key members of the non-canonical Wnt/Ca2+ signaling pathway is also detected following PFN1 suppression, providing the groundwork for future studies, addressing the specific role of PFN1 in Ca2+ signaling, particularly in the muscle invasive disease.


Creative Commons License All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 License.
PII: 12218