Enzalutamide inhibits proliferation of gemcitabine-resistant bladder cancer cells with increased androgen receptor expression

Kameyama,Koji; Horie,Kengo; Mizutani,Kosuke; Kato,Taku; Fujita,Yasunori; Kawakami,Kyojiro; Kojima,Toshio; Miyazaki,Tatsuhiko; Deguchi,Takashi; Ito,Masafumi

doi:10.3892/ijo.2016.3781

Enzalutamide inhibits proliferation of gemcitabine-resistant bladder cancer cells with increased androgen receptor expression

Authors:
- Koji Kameyama
- Kengo Horie
- Kosuke Mizutani
- Taku Kato
- Yasunori Fujita
- Kyojiro Kawakami
- Toshio Kojima
- Tatsuhiko Miyazaki
- Takashi Deguchi
- Masafumi Ito
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Affiliations: Department of Urology, Gifu University Graduate School of Medicine, Gifu, Gifu 501-1193, Japan, Research Team for Mechanism of Aging, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173-0015, Japan, Health Support Center, Toyohashi University of Technology, Tenpaku-cho, Toyohashi, Aichi 441-8580, Japan, Division of Pathology, Gifu University Hospital, Gifu, Gifu 501-1194, Japan
Published online on: November 24, 2016 https://doi.org/10.3892/ijo.2016.3781
Pages: 75-84

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Abstract

Advanced bladder cancer is treated mainly with gemcitabine and cisplatin, but most patients eventually become resistance. Androgen receptor (AR) signaling has been implicated in bladder cancer as well as other types of cancer including prostate cancer. In this study, we investigated the expression and role of AR in gemcitabine-resistant bladder cancer cells and also the potential of enzalutamide, an AR inhibitor, as a therapeutic for the chemoresistance. First of all, we established gemcitabine-resistant T24 cells (T24GR) from T24 bladder cancer cells and performed gene expression profiling. Microarray analysis revealed upregulation of AR expression in T24GR cells compared with T24 cells. AR mRNA and protein expression was confirmed to be increased in T24GR cells, respectively, by quantitative RT-PCR and western blot analysis, which was associated with more potent AR transcriptional activity as measured by luciferase reporter assay. The copy number of AR gene in T24GR cells determined by PCR was twice as many as that of T24 cells. AR silencing by siRNA transfection resulted in inhibition of proliferation of T24GR cells. Cell culture in charcoal-stripped serum and treatment with enzalutamide inhibited growth of T24GR cells, which was accompanied by cell cycle arrest. AR transcriptional activity was found to be reduced in T24GR cells by enzalutamide treatment. Lastly, enzalutamide also inhibited cell proliferation of HTB5 bladder cancer cells that express AR and possess intrinsic resistance to gemcitabine. Our results suggest that enzalutamide may have the potential to treat patients with advanced gemcitabine-resistant bladder cancer with increased AR expression.

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