For example, Wnt signalling has always enhanced CSC properties,39 Toll-like receptor signalling facilitates stem cell marker expression in HCC,40 and the PI3K-AKT pathway is a canonical signalling pathway downstream of EGFR to reduce tumour-initiating cell frequency.41 The Hippo-YAP pathway showed the greatest upregulation in response to CBX4 in our results. This evidence concerns the gene AKT1 and hepatocellular carcinoma.