Received March 30, 2016; Accepted August 22, 2016; Published December 26, 2016
http://dx.doi.org/10.18081/2333-5106/016-314-328
Joan V. Draucker; Irene Talarico; Ling Wu; David B. Mulier; Xian Chen; Celeste B. Determan; Jean Friese; Ling Yao
Abstract
Bladder carcinoma is the most common malignancy of the urinary system involving bladder epithelium. According to the Global Cancer Observatory, it has an age-adjusted incidence rate of 7.1 per 100,000 men per year. In transitional cell bladder carcinoma T24 cells, Notch-3 was established as a promoter of tumor growth through reprogramming the metabolic active state. Transcriptomic profiling and follow-up analyses confirmed the activities of oncogenic signals, including AMPK repression and HIF-1α pathway incorporation. Particularly, for the fraction of HIF-1α target genes related to fatty acid biosynthesis, their roles in promoting proliferation and anchorage independence were further demonstrated. Mechanistically, Notch-3 was shown to stabilize HIF-1α protein in an intracellular vesicular compartment, thereby boosting the transcriptional activity of HIF-1α downstream targets. The vesicular sequestration was attributed to the promotion of endosomal acidification, a process relying on Notch-3-mediated Rab14 activation. The significance of Notch-3 signaling in promoting carcinogenesis and tumor progression was corroborated by gain- and loss-of-function approaches in orthotopic cancer models. Overall, Notch-3 reprograms the metabolic state and promotes tumorigenesis by activating the HIF-1α signaling pathway. Aberrant Notch signaling has been implicated in various human malignancies. In bladder carcinoma, however, the expression levels, tumor-promoting or suppressing activities, and the underlying mechanisms of Notch receptors have not been well characterized. Transcriptional signatures of each Notch receptor were determined in T24 and UM-UC-3 cells. Stably transfected T24 cells expressing Notch-3 construct or a dominant-negative Notch-3 mutant (akin to Notch-3KD) were developed. Shunt transfectants against Notch-3 and Notch-3 pseudogenic were established. Tumor and vesicle tissue lysates were prepared. Transcriptomic profiling was performed using microarray and qPCR analyses.
Keyword: Bladder cancer; Noch-3; PI3K/Akt pathway; Angiogenesis; Proliferation
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