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The Scientific World Journal
Volume 2013, Article ID 174392, 7 pages
http://dx.doi.org/10.1155/2013/174392
Research Article

Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells

1Eulji Medi-Bio Research Institute, Seoul 139-871, Republic of Korea
2Department of Urology, College of Medicine, Eulji University, Hangeulbiseok-gil, Hagye-dong, Nowon-ku, Seoul 139-871, Republic of Korea
3Department of Biotechnology, Seoul Women’s University, Seoul 139-871, Republic of Korea

Received 22 April 2013; Accepted 22 May 2013

Academic Editors: A. Tefekli, V. Tugcu, and M. Tunc

Copyright © 2013 Sang Soo Kim et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction in any medium, provided the original work is properly cited.

Abstract

We evaluated effect of dual gene silencing of Hsp27 and c-FLIP in doxazosin-induced apoptosis of PC-3 cell. After transfection using Hsp27 and c-FLIP siRNA mixture (dual silencing), doxazosin treatment was done at the concentrations of 1, 10, and 25 μM. We checked apoptosis of PC-3 cells with and TUNEL staining. We also checked interaction between Hsp27 and C-FLIP in the process of apoptosis inhibition. Spontaneous apoptotic index was 5% under single gene silencing of Hsp27 and c-FLIP and 7% under dual silencing of Hsp27 and c-FLIP. When doxazosin treatment was added, apoptotic indices increased in a dose-dependent manner (1, 10, and 25 μM): nonsilencing 10, 27, and 52%; Hsp27-silencing: 14, 35, and 68%; c-FLIP silencing: 21, 46, and 78%; dual silencing: 38, 76, and 92%. While c-FLIP gene expression decreased in Hsp27- silenced cells, Hsp27 gene expression showed markedly decreased pattern in the cells of c-FLIP silencing. The knockout of c-FLIP and Hsp27 genes together enhances apoptosis even under 1 μM, rather than low concentration, of doxazosin in PC-3 cells. This finding suggests a new strategy of multiple knockout of antiapoptotic and survival factors in the treatment of late-stage prostate cancer refractory to conventional therapy.