Esculentoside A suppresses the malignant progression of hepatocellular carcinoma by promoting RORB.

Hepatocellular carcinoma (HCC) is a highly aggressive disease associated with a poor prognosis. The present study attempts to discover the potential of esculentoside A (EsA) in inhibiting HCC progression. Cell viability was assessed using the Cell Counting Kit-8 assays. At the same time, stemness and malignant behaviors were evaluated through sphere formation, Western blot (WB), flow cytometry, colony formation assays, and TUNEL. EsA treatment suppressed HCC cell proliferation, migration, invasion, and stemness in vitro and tumor growth in vivo. Bioinformatics identified retinoic acid receptor-related orphan receptor beta (RORB) as a potential target of EsA, and EsA induced RORB expression in HCC cells. EsA also inhibited Wnt signaling activity, as shown by TOP/FOP Flash assays and WB. Knockdown of RORB through lentivirus infection reversed the effects of EsA. At the same time, Wnt pathway inhibitor rescued the promotive role of RORB knockdown in HCC malignant behaviors, suggesting that EsA inhibited HCC progression via the RORB/Wnt axis. RORB expression was reduced and associated with higher Wnt signaling in tumor tissues from patients with HCC, and low RORB expression showed significant correlation with higher TNM staging in HCC patients. This study offers novel insights into the molecular mechanism of EsA in HCC.