BACKGROUND & AIMS: The Notch signaling pathway is activated in leukemia and 
solid tumors (such as lung cancer), but little is known about its role in liver 
cancer.
METHODS: The intracellular domain of Notch was conditionally expressed in 
hepatoblasts and their progeny (hepatocytes and cholangiocytes) in mice. This 
was achieved through Cre expression under the control of an albumin and 
α-fetoprotein (AFP) enhancer and promoter (AFP-Notch intracellular domain 
[NICD]). We used comparative functional genomics to integrate transcriptome data 
from AFP-NICD mice and human hepatocellular carcinoma (HCC) samples (n = 683). A 
Notch gene signature was generated using the nearest template prediction method.
RESULTS: AFP-NICD mice developed HCC with 100% penetrance when they were 12 
months old. Activation of Notch signaling correlated with activation of 3 
promoters of insulin-like growth factor 2; these processes appeared to 
contribute to hepatocarcinogenesis. Comparative functional genomic analysis 
identified a signature of Notch activation in 30% of HCC samples from patients. 
These samples had altered expression in Notch pathway genes and activation of 
insulin-like growth factor signaling, despite a low frequency of mutations in 
regions of NOTCH1 associated with cancer. Blocking Notch signaling in liver 
cancer cells with the Notch activation signature using γ-secretase inhibitors or 
by expressing a dominant negative form of mastermind-like 1 reduced their 
proliferation in vitro.
CONCLUSIONS: Notch signaling is activated in human HCC samples and promotes 
formation of liver tumors in mice. The Notch signature is a biomarker of 
response to Notch inhibition in vitro.