Comprehensive Molecular Profiling of Combined Hepatocellular Carcinoma and Cholangiocarcinoma reveals Distinct Notch Signaling Subgroups with Prognostic Significance

Combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CCC) exhibits distinct Notch signaling activation patterns, with elevated Notch1 and downstream target expression compared to HCC and CCC. Stratification into three subgroups based on Notch1 and Hes5 expression revealed that Notch-responsive tumors (high Notch1 and Hes5) demonstrated aggressive biological behavior, poor prognosis, and molecular profiles resembling CCC. These findings underscore the heterogeneity of cHCC-CCC and suggest that Notch1 and Hes5 serve as potential biomarkers for risk stratification and therapeutic targeting, particularly for Notch pathway and immune checkpoint inhibitors. This study involved the molecular profiling of combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CCC), hepatocellular carcinoma (HCC), and cholangiocarcinoma (CCC). RNA sequencing and immunohistochemical staining were performed to assess Notch signaling activation. Samples were categorized into three subgroups based on Notch1 and Hes5 expression levels.