Arsenic trioxide and sorafenib combination therapy for human hepatocellular carcinoma functions via up-regulation of TNF-related apoptosis-inducing ligand.

Lingyan Wang,Zhihui Min,Yanhong Wang,Zhenggang Ren,Dongli Song,Xiangdong Wang,Yunfeng Cheng,Mushuang Hu

doi:10.3892/ol.2018.8981

Abstract

The survival benefits of sorafenib treatment for patients with hepatocellular carcinoma (HCC) are limited due to drug resistance and side effects. Therefore, combinations of sorafenib with other low toxicity drugs, including arsenic trioxide (As2O3) require investigation. The present study aimed to evaluate the potency of apoptosis-induction by As2O3/sorafenib treatment in HCC cell lines, Huh7, 97H and freshly-isolated HCC cells, and also to elucidate the underlying mechanism. A total of 10 patients with HCC were enrolled in the present study. Freshly-isolated HCC cells were purified from HCC tissues collected at surgery. HCC-cell apoptosis was measured by flow cytometry using proprium iodide/Annexin-V staining. The impacts of As2O3 and/or sorafenib on Huh7, 97H and fresh-isolated HCC-cell proliferation were evaluated by Cell Counting Kit-8 assay. The expression of TNF-related apoptosis-inducing ligand (TRAIL) was determined by reverse transcription-quantitative polymerase chain reaction and western blotting. The downregulation of TRAIL protein expression was achieved using small interfering RNA. The combination of As2O3 and sorafenib had anti-proliferative and pro-apoptotic effects in the liver cancer cell line, Huh7, via increased expression of TRAIL, but not in 97H cells. TRAIL-knockdown increased the drug-resistance of Huh7 cells. Freshly-isolated HCC cells were more sensitive to the As2O3 and sorafenib combination than the single drug treatments. Overall, the combination of As2O3 and sorafenib demonstrated potent anti-tumor activity in Huh7 and freshly-isolated HCC cells via a TRAIL-dependent pathway. This may be a potential therapeutic approach for advanced HCC treatment.

Highlights

To provide a novel solution to sorafenib‐resistance in hepatocellular carcinoma (HCC), the present study aimed to evaluate the efficacy of arsenic trioxide (As2O3)/sorafenib combined treatment against primary HCC cells and its impact on TNF‐related apoptosis‐inducing ligand (TRAIL) expression
To investigate the effect of As2O3 with and without sorafenib on liver cancer cell lines, Huh7 and 97H, cells were stimulated with drugs at different concentrations for 72 h
Huh7‐cell proliferation was inhibited in an As2O3‐concnetration‐dependent manner when treated with sorafenib in the presence of varying concentrations of As2O3 (Fig. 1)

Summary

Introduction

As the second leading cause of cancer‐associated mortality in men, and the sixth in women worldwide [1], human hepatocellular carcinoma (HCC) remains a severe global health threat. Chronic infections of hepatitis B virus (HBV) and hepatitis C virus (HCV) are major risk factors associated with HCC [2]. With an increased rate of HBV/HCV infection and other risk factors, including obesity, cigarette smoking, diabetes mellitus and heavy alcohol consumption, the incidence of HCC has increased [3]. In order to prevent HCC progression, tumor resection, liver transplantation and radiofrequency (thermal) ablation [RF(T)A] are commonly used for HCC treatment [4,5,6]. These treatments have improved the survival rates of patients with HCC, they are only appropriate for a minority of patients, and their efficacies are limited by high recurrence rates [7].

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Results

Conclusion