Abstract
Despite comprehensive genomic analyses, no targeted therapies are approved for bladder cancer. Here, we investigate whether a single and combination therapy with targeted agents exert antitumor effects on bladder cancer cells through genomic alterations using a three-dimensional (3D) high-throughput screening (HTS) platform. Seven human bladder cancer cell lines were used to screen 24 targeted agents. The effects of 24 targeted agents were dramatically different according to the genomic alterations of bladder cancer cells. BEZ235 (dual phosphatidylinositol-3-kinase (PI3K)/mammalian target of rapamycin (mTOR) inhibitor) showed antitumor effects against most cell lines, while AZD2014 (mTOR inhibitor) had an IC50 value lower than 2 μM in 5637, J82, and RT4 cell lines. AZD5363 (protein kinase B (AKT) inhibitor) exerted antitumor effects on 5637, J82, and 253J-BV cells. J82 cells (PI3KCA and mTOR mutations) were sensitive to AZD5363, AZD2014, and BEZ235 alone or in AZD5363/AZD2014 and AZD5363/BEZ235 combinations. Although all single drugs suppressed cell proliferation, the combination of drugs exhibited synergistic effects on cell viability and colony formation. The synergistic effects of the combination therapy on the PI3K/Akt/mTOR pathway, apoptosis, and EMT were evident in Western blotting. Thus, the 3D culture-based HTS platform could serve as a useful preclinical tool to evaluate various drug combinations.
Highlights
Bladder cancer is the most common malignancy of the urinary tract with 430,000 newly diagnosed cases in 2012
The phosphoinositide 3 kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin pathway is the key driver of carcinogenesis and progression in bladder cancer [9]
We focused on the effect of AZD5363, AZD2014, and BEZ235 alone or in combination on two bladder cancer cell lines differing in their mammalian target of rapamycin (mTOR) mutation status
Summary
Bladder cancer is the most common malignancy of the urinary tract with 430,000 newly diagnosed cases in 2012. The phosphoinositide 3 kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway is the key driver of carcinogenesis and progression in bladder cancer [9]. The PI3K/AKT/mTOR pathway is a potential therapeutic target for bladder cancer. We investigated whether different targeted agents exert antitumor effects against bladder cancer cells, through genomic alterations, using a three-dimensional (3D) high-throughput screening (HTS) platform. We focused on the effect of AZD5363 (an AKT inhibitor), AZD2014 (an mTOR inhibitor), and BEZ235 (a dual PI3K/mTOR inhibitor) alone or in combination on two bladder cancer cell lines differing in their mTOR mutation status. We assessed the expression patterns of the markers of the PI3K/AKT/mTOR pathway (p-AKT, p-mTOR, p-S6), apoptosis (poly (ADP-ribose) polymerase (PARP), cleaved caspase-3, B cell lymphoma-2 (Bcl-2), Bax), and epithelial-to-mesenchymal transition (EMT) (phosphor-signal transducer and activator of transcription 3 (p-STAT3), E-cadherin, snail, vimentin) by Western blotting
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
