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

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 TU...

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Main Authors: Sang Soo Kim, Hee-Ju Cho, Jeong-Man Cho, Jung Yoon Kang, Hyun-Won Yang, Tag Keun Yoo
Format: Article
Language:English
Published: Wiley 2013-01-01
Series:The Scientific World Journal
Online Access:http://dx.doi.org/10.1155/2013/174392
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author Sang Soo Kim
Hee-Ju Cho
Jeong-Man Cho
Jung Yoon Kang
Hyun-Won Yang
Tag Keun Yoo
author_facet Sang Soo Kim
Hee-Ju Cho
Jeong-Man Cho
Jung Yoon Kang
Hyun-Won Yang
Tag Keun Yoo
author_sort Sang Soo Kim
collection DOAJ
description 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.
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spelling doaj-art-5de464003db34c0a9e3352c41fa8c1762025-08-20T02:06:03ZengWileyThe Scientific World Journal1537-744X2013-01-01201310.1155/2013/174392174392Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer CellsSang Soo Kim0Hee-Ju Cho1Jeong-Man Cho2Jung Yoon Kang3Hyun-Won Yang4Tag Keun Yoo5Eulji Medi-Bio Research Institute, Seoul 139-871, Republic of KoreaDepartment of Urology, College of Medicine, Eulji University, Hangeulbiseok-gil, Hagye-dong, Nowon-ku, Seoul 139-871, Republic of KoreaDepartment of Urology, College of Medicine, Eulji University, Hangeulbiseok-gil, Hagye-dong, Nowon-ku, Seoul 139-871, Republic of KoreaDepartment of Urology, College of Medicine, Eulji University, Hangeulbiseok-gil, Hagye-dong, Nowon-ku, Seoul 139-871, Republic of KoreaDepartment of Biotechnology, Seoul Women’s University, Seoul 139-871, Republic of KoreaEulji Medi-Bio Research Institute, Seoul 139-871, Republic of KoreaWe 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.http://dx.doi.org/10.1155/2013/174392
spellingShingle Sang Soo Kim
Hee-Ju Cho
Jeong-Man Cho
Jung Yoon Kang
Hyun-Won Yang
Tag Keun Yoo
Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells
The Scientific World Journal
title Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells
title_full Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells
title_fullStr Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells
title_full_unstemmed Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells
title_short Dual Silencing of Hsp27 and c-FLIP Enhances Doxazosin-Induced Apoptosis in PC-3 Prostate Cancer Cells
title_sort dual silencing of hsp27 and c flip enhances doxazosin induced apoptosis in pc 3 prostate cancer cells
url http://dx.doi.org/10.1155/2013/174392
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AT jungyoonkang dualsilencingofhsp27andcflipenhancesdoxazosininducedapoptosisinpc3prostatecancercells
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