Berberine Sensitizes Human Hepatoma Cells to Regorafenib via Modulating Expression of Circular RNAs.

Kunyuan Wang,Jiaen Lin,Zhilei Wang,Chengxin Gu,Hui Yang,Tao Yang,Shiming Liu,Qianting Lu,Ganxiang Yu

doi:10.3389/fphar.2021.632201

Abstract

Regorafenib resistance is a key limiting factor in the treatment of advanced hepatocellular carcinoma (HCC). Increasing evidence has demonstrated that Berberine (BBR) can synergistically enhance the therapeutic effect of various chemotherapeutic agents. However, the contribution of BBR on regorafenib therapy remains unclear. The purpose of this study was to explore the combined treatment effect of berberine and regorafenib in HCC. We found that BBR enhanced the cytotoxicity of regorafenib in HCC cells. Compared with regorafenib alone, the combined treatment of BBR and regorafenib significantly inhibited the proliferation of HCC cells and induced cellular apoptosis. Meanwhile, the combined treatment group with BBR (10mg/kg/day) and regorafenib (5mg/kg/day) had a dramatic inhibitory effect on the growth of HCC xenograft tumors in nude mice. The increased apoptosis of xenograft tumors was seen in the combined treatment group. Moreover, a comprehensive circular RNA sequencing was performed to identify differentially expressed circRNAs in HCC cells after exposure to 100µM BBR and 5µM regorafenib. The volcano plot and scatter plot analyses revealed that there were 58 up-regulated and 19 down-regulated differentially expressed circRNAs between the combination treatment and control groups. Among them, the expression of hsa_circ_0032029 and hsa_circ_0008928 were up-regulated in HCC cells after treatment with 100µM BBR and 5µM regorafenib. Taken together, this study demonstrated that BBR enhanced the anti-HCC effect of regorafenib both in vitro and in vivo. The synergistic anti-tumor effect of BBR and regorafenib might be related to the up-regulation of hsa_circ_0032029 and hsa_circ_0008928 in HCC cells.

Highlights

Hepatocellular carcinoma (HCC) is a major health problem with increasing incidence and mortality (Bray et al, 2018; Forner et al, 2018)
To investigate cellular apoptosis induced by BBR and regorafenib, hepatocellular carcinoma (HCC) cells were treated with BBR and regorafenib alone or in combination
We found that the combined treatment of BBR and regorafenib significantly induced HCC cells apoptosis when cells were cultured over 48h (Figure 3A)

Summary

Introduction

Hepatocellular carcinoma (HCC) is a major health problem with increasing incidence and mortality (Bray et al, 2018; Forner et al, 2018). Regorafenib, an oral multi-kinase inhibitor, has become a therapeutic agent which significantly improves overall survival after the treatment failure of sorafenib, and receives the Berberine Enhances Regorafenib’ Anti-HCC Effect approval as a second line treatment for advanced HCC in 2017 (Bruix et al, 2017; Duffy and Greten, 2017). It is needed to find novel agents that can increase the sensitivity of HCC cells to regorafenib treatment. Our previous study has proved that BBR makes HCC cells sensitive to sorafenib (Huang et al, 2018). It is unclear whether BBR can enhance the anti-HCC activity of regorafenib

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Results

Conclusion