Simultaneous silencing of ACSL4 and induction of GADD45B in hepatocellular carcinoma cells amplifies the synergistic therapeutic effect of aspirin and sorafenib

Hongping Xia,Amudha Deivasigamani,Veerabrahma Pratap Seshachalam,Brian Kim Poh Goh,Jianxiang Chen,Kee Wah Lee,London Lucien Ooi,Kam M Hui,Karthik Sekar,Shik Nie Kong

doi:10.1038/cddiscovery.2017.58

Abstract

Sorafenib is currently the only US Food and Drug Administration (FDA)-approved molecular inhibitor for the systemic therapy of advanced hepatocellular carcinoma (HCC). Aspirin has been studied extensively as an anti-inflammation, cancer preventive and therapeutic agent. However, the potential synergistic therapeutic effects of sorafenib and aspirin on advanced HCC treatment have not been well studied. Drug combination studies and their synergy quantification were performed using the combination index method of Chou-Talalay. The synergistic therapeutic effects of sorafenib and aspirin were evaluated using an orthotopic mouse model of HCC and comprehensive gene profiling analyses were conducted to identify key factors mediating the synergistic therapeutic effects of sorafenib and aspirin. Sorafenib was determined to act synergistically on HCC cells with aspirin in vitro. Using Hep3B and HuH7 HCC cells, it was demonstrated that sorafenib and aspirin acted synergistically to induce apoptosis. Mechanistic studies demonstrated that combining sorafenib and aspirin yielded significant synergistically anti-tumor effects by simultaneously silencing ACSL4 and the induction of GADD45B expression in HCC cells both in vitro and in the orthotopic HCC xenograft mouse model. Importantly, clinical evidence has independently corroborated that survival of HCC patients expressing ACSL4highGADD45Blow was significantly poorer compared to patients with ACSL4lowGADD45Bhigh, thus demonstrating the potential clinical value of combining aspirin and sorafenib for HCC patients expressing ACSL4highGADD45Blow. In conclusion, sorafenib and aspirin provide synergistic therapeutic effects on HCC cells that are achieved through simultaneous silencing of ACSL4 and induction of GADD45B expression. Targeting HCC with ACSL4highGADD45Blow expression with aspirin and sorafenib could provide potential synergistic therapeutic benefits.

Highlights

Being a clear example of inflammation-driven cancer,[9] we have explored the possibility of aspirin to enhance the therapeutic effects of sorafenib in a preclinical model of advanced orthotopic Hepatocellular carcinoma (HCC)
To investigate the anticancer therapeutic effects of aspirin and sorafenib, we first employed a panel of liver cancer cell lines including Hep3B, HLE, HuH7, SK-HEP-1 and HCCLM3 to assess their growth inhibition response to aspirin and sorafenib treatment
For patients with unresectable or metastatic HCC, conventional chemotherapy is of limited benefit

Summary

Introduction

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths worldwide and the global incidence is rising.An estimated 782 500 new liver cancer cases and 745 500 deaths occurred worldwide during 2012, with China alone accounting for about 50% of the total number of cases and deaths.[1,2] most HCC patients are still being diagnosed in a late stage and the only treatment available to these patients is sorafenib.[3] the data from the Sorafenib Hepatocellular Carcinoma Assessment Randomized Protocol (SHARP) trial[4] and the Asia-Pacific study[5] could demonstrate a significant survival benefit, the absolute gain in life expectancy was marginal (2–3 months overall survival benefit compared with placebo) and the cost effectiveness of sorafenib is a big concern. Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths worldwide and the global incidence is rising. An estimated 782 500 new liver cancer cases and 745 500 deaths occurred worldwide during 2012, with China alone accounting for about 50% of the total number of cases and deaths.[1,2] most HCC patients are still being diagnosed in a late stage and the only treatment available to these patients is sorafenib.[3]. It has recently been shown to protect against certain types of cancer.[8] Being a clear example of inflammation-driven cancer,[9] we have explored the possibility of aspirin to enhance the therapeutic effects of sorafenib in a preclinical model of advanced orthotopic HCC

Methods

Results

Conclusion