Abstract 2546: Targeting of AKT/β-catenin signaling pathways driven hepatocellular carcinoma cell lines by combined treatment of HDAC and multi-kinase inhibitors

Abstract

Abstract Hepatocellular carcinoma (HCC) is the dominant type of liver cancer exhibiting high mortality and poor prognosis with the fastest rate of death due to its aggressiveness and lack of effective therapies. Beta-catenin is a key player transcriptional factor, which orchestrates gene expression in HCC progression. Activation of β-catenin is promoted by its phosphorylation by AKT. Although the studies of metastatic behavior of β-catenin over-expressing HCC cells are not clear, the present study is focusing on the relationship between chromatin remodelling through deacetylation and phosphorylation of β-catenin by AKT. The current study investigated the role of a combination of Lenvatinib, a multi-kinase inhibitor, and Belinostat, a histone deacetylase inhibitor (HDACI), on two HCC cell lines, Hep G2 and Hep 3B, respectively. In-depth, our data showed that combined treatment significantly induced morphological changes in the tested cell lines with activation of caspase-3 and pre-apoptotic signals of Bcl-2, Bcl-XL, and Bax, indicating apoptotic induction after 48 hours of treatment compared to the control or each drug separately. The calculated IC50 was 1µmol/L for Belinostat and 25 n mole/L for Lenvatinib. Western blotting analysis reveals that the combined treatment for 48 hours inhibited the oncogenic β-catenin, AKT, and TGFβ expression in a dose-dependent manner.In addition, the combined treatment inhibited the localization of beta-catenin in the tested cell nuclei after 48 hours as confirmed by IHC staining. Statistical analysis proved that the combination of both drugs significantly decreased the proliferation and invasion of both cell lines (P<0.01, ANOVA) compared to each drug alone. In addition, the percentage of apoptotic cells was significantly increased and showed a cell cycle arrest in the G0/G1 phase of Hep-G2 cells after 48 hours of combined treatment as confirmed by flow cytometry. All in all, our data show that targeting AKT/β-catenin driven HCC progression can be targeted by Lenvatinib and Belinostat combined treatment. Citation Format: Mohamed Mahmoud Elberawi, Mohamed Abdulqadir, Ahmed Samir Sultan. Targeting of AKT/β-catenin signaling pathways driven hepatocellular carcinoma cell lines by combined treatment of HDAC and multi-kinase inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2546.