1B and 4B). This finding clearly indicates that a growth-suppressive environment was generated in hyperplastic livers, preventing their further growth. On the other hand, the development of HCC in all mice given the genotoxic agent DENA prior to TCPOBOP suggests that initiated/mutated cells have escaped the growth-suppressive signals, thus clonally expanding to develop HCC. Our finding of increased YAP protein expression and its nuclear translocation see more in HCC cells suggests that dysregulation of the Hippo pathway may contribute to the

escape from the environmental growth-suppressive constraint; it is noteworthy that a strong and increased nuclear YAP staining has been observed in human tumors, including HCC.27, 30 In this study, we show that YAP nuclear translocation is accompanied by its increased activity because, in the same tumors, up-regulation of AFP and CTGF, two YAP target genes,15, 17 was observed. Taken

together, our findings suggest that YAP dysregulation could be involved in the development of DENA+TCPOBOP-induced HCC. MicroRNAs have recently emerged as important modulators of gene expression in cancer,31 including human HCC.32 Very recently, Liu et al.29 reported that miR-375 is a negative regulator of YAP; indeed, they found down-regulation Cabozantinib purchase of miR-375 in tumor tissues of HCC patients, which was accompanied by increased YAP levels. Moreover, they showed that miRNA-375 re-expression caused a severe decrease of YAP protein levels. In accordance with these results, we found a decrease of miR-375 and an increase

in YAP content in approximately 70% of mouse HCCs. Our data thus MCE provide a possible mechanism underlying the increase of YAP in chemically induced HCCs. Whether down-regulation of miR-375 is due to epigenetic modifications is presently unknown and warrants further investigation, because modulation of this microRNA could be therapeutically targeted to reactivate the growth-suppressive effect of the Hippo pathway. A better understanding of growth regulatory mechanisms may represent an important approach from a therapeutic point of view. HCC, the fifth most common malignant neoplasm and the third most frequent cause of cancer-related death worldwide, represents a major health problem.33, 34 A better definition of the molecular pathogenesis of HCC could have a significant impact on the development of new treatment strategies. The Hippo kinase cascade might have clear pathogenic implications in hepatocarcinogenesis, and its drivers might represent novel targets for molecular therapies.