Urology, National University Health System, 5 Lower Kent Ridge Road, Singapore, 119074, Singapore.
We investigated the effect of the mTOR inhibitor everolimus (RAD001) on human bladder cancer (BC) cells in vitro and in vivo.
The effect of RAD001 on the growth of UM-UC-3, UM-UC-6, UM-UC-9, and UM-UC-14 BC cells were assessed by crystal violet and [3H]Thymidine incorporation assays . Flow cytometry cell cycle analyses were performed to measure the apoptotic cell fraction. Protein synthesis was measured using tritium-labeled leucine-incorporation assays. The effects of RAD001 on the mTOR pathway were analyzed by western blotting. To test the effects of RAD001 in vivo, UM-UC-3, UM-UC-6, and UM-UC-9 cells were subcutaneously implanted into nude mice. Tumor-bearing mice were treated orally with RAD001 or placebo. Tumors were harvested for immunohistochemical analysis.
In vitro, RAD001 transiently inhibited BC cell growth in a dose-dependent manner. This effect was augmented by retreatment of cells after 3 days. UM-UC-14 cells were the most sensitive to RAD001, while UM-UC-9 cells were the least sensitive. After retreatment with RAD001, only sensitive cell lines showed G1 phase arrest, with no evidence of apoptosis. RAD001 significantly inhibited the growth of tumors that were subcutaneously implanted in mice. Inhibition of protein synthesis through the S6K and 4E-BP1 pathways appears to be the main mechanism for the RAD001-induced growth inhibition. However, inhibition of angiogenesis was the predominant mechanism of RAD001's effect on UM-UC-9 cells.
The mTOR inhibitor RAD001 inhibits growth of BC cells in vitro. RAD001 is effective in treating BC tumors in an in vivo nude mouse model despite the heterogeneity of in vitro responses.
Written by:
Chiong E, Lee IL, Dadbin A, Sabichi AL, Harris LD, Urbauer DL, McConkey DJ, Dickstein RJ, Cheng T, Grossman HB. Are you the author?
Reference: Clin Cancer Res. 2011 Mar 17. Epub ahead of print.
doi: 10.1158/1078-0432.CCR-09-3202
PubMed Abstract
PMID: 21415218
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