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Asian Journal of Andrology (2009) 11: 119-126. doi: 10.1038/aja.2008.26; published online 15 December 2008.
Chimeric molecules facilitate the degradation of androgen receptors and repress the growth of LNCaP cells
Yue-Qing Tang1,2, Bang-Min Han1, Xin-Quan Yao3, Yan Hong1,2, Yan Wang4, Fu-Jun Zhao1, Sheng-Qiang Yu1,2, Xiao-Wen Sun1 and Shu-Jie Xia1,2
1 Department of Urology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200025, China
2 Institute of Urology, Shanghai Jiao Tong University, Shanghai 200025, China
3 Department of Urology, Wujiang Third People's Hospital, Suzhou 215228, China
4 Department of Immunology, Shanghai Jiao Tong University, Shanghai 200025, China
Correspondence: Prof. Shu-Jie Xia, Department of Urology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China. Fax:+86-21-6324-1377 E-mail: xsjurologist@163.com; Dr Xiao-Wen Sun, Department of Urology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China. Fax:+86-21-6324-1377 E-mail: sunxiaow1973@163.com
Received 8 October 2008; Accepted 13 October 2008; Published online 15 December 2008.
| Abstract |
Post-translational degradation of protein plays an important role in cell life. We employed chimeric molecules (dihydrotestosterone-based proteolysis-targeting chimeric molecule [DHT-PROTAC]) to facilitate androgen receptor (AR) degradation via the ubiquitin?proteasome pathway (UPP) and to investigate the role of AR in cell proliferation and viability in androgen-sensitive prostate cancer cells. Western blot analysis and immunohistochemistry were applied to analyse AR levels in LNCaP cells after DHT-PROTAC treatment. Cell counting and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cell viability assay were used to evaluate cell proliferation and viability after AR elimination in both LNCaP and PC-3 cells. AR was tagged for elimination via the UPP by DHT-PROTAC, and this could be blocked by proteasome inhibitors. Degradation of AR depended on DHT-PROTAC concentration, and either DHT or an ALAPYIP-(arg)8 peptide could compete with DHT-PROTAC. Inhibition of cell proliferation and decreased viability were observed in LNCaP cells, but not in PC-3 or 786-O cells after DHT-PROTAC treatment. These data indicate that AR elimination is facilitated via the UPP by DHT-PROTAC, and that the growth of LNCaP cells is repressed after AR degradation.
Keywords: androgen receptor, LNCaP, prostate cancer, proteolysis, ubiquitin |
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