Abstract
Activating mutation of the fibroblast growth factor receptor-3 (FGFR3) gene is known as a key molecular event in both oncogenesis and cell proliferation of low-grade noninvasive human bladder urothelial carcinoma (UC), which is characterized by frequent intravesical recurrence. In this study, we investigated the antitumor potentiality of 1-tert-butyl-3-[6-(3,5-dimethoxy-phenyl)-2-(4-diethylamino-butylamino)-pyrido[2,3-d]pyrimidin-7-yl]-urea (PD173074), a small-molecule FGFR3-selective tyrosine kinase inhibitor (TKI), as a therapeutic modality using eight UC cell lines. In our in vitro cell proliferation assay, PD173074 suppressed cell proliferation remarkably in two cell lines, namely, UM-UC-14 and MGHU3, which expressed mutated FGFR3 protein. In contrast, the other six cell lines expressing wild-type FGFR3 or without FGFR3 expression were resistant to PD173074 treatment. Cell cycle analysis revealed the growth inhibitory effect of PD173074 was associated with arrest at G1-S transition in a dose-depending manner. Furthermore, we observed an inverse relationship between Ki-67 and p27/Kip1 expression after PD173074 treatment, suggesting that up-regulation of p27 recruited UC cells harboring activating FGFR3 mutations in G1 that was analogous with the other receptor TKIs acting on the epidermal growth factor receptors. In the mouse xenograft models using subcutaneously transplanted UM-UC-14 and MGHU3, orally administered PD173074 suppressed tumor growth and induced apoptotic changes comparable with the results of our in vitro assay. These findings elucidated the effectiveness of molecular targeted approach for bladder UC harboring FGFR3 mutations and the potential utility to decrease the intravesical recurrence of nonmuscle invasive bladder UC after transurethral surgical resection.
Footnotes
This work was supported in part by the Ministry of Health, Labor and Welfare, Japan [Grants-in-Aid for Cancer Research G21S-9-2] (to K.S.); and the 3rd Term Comprehensive 10-Year Strategy for Cancer Control [Grant 19-007] (to K.S.).
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.109.162768.
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ABBREVIATIONS:
- UC
- urothelial carcinoma
- FGFR
- fibroblast growth factor receptor
- PD173074
- 1-tert-butyl-3-[6-(3,5-dimethoxy-phenyl)-2-(4-diethylamino-butylamino)-pyrido[2,3-d]pyrimidin-7-yl]-urea
- TKI
- tyrosine kinase inhibitor
- SU5402
- 3-((3-(2-carboxyethyl)-4-methylpyrrol-2-yl) methylene)-2-inolinone
- EGFR
- epidermal growth factor receptor
- aFGF
- acidic fibroblast growth factor
- IHC
- immunohistochemical staining
- H&E
- hematoxylin and eosin
- PI3K
- phosphatidylinositol 3-kinase
- CHIR-258
- 4-amino-5-fluoro-3-[6-(4-methyl-1-piperazinyl)-1H-benzimidazol-2-yl]-2(1H-quinolinone.
- Received October 20, 2009.
- Accepted December 1, 2009.
- Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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