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Open Access Research article

Differential effects of arsenic trioxide on chemosensitization in human hepatic tumor and stellate cell lines

Fatima Rangwala1, Kevin P Williams2, Ginger R Smith2, Zainab Thomas2, Jennifer L Allensworth3, H Kim Lyerly34, Anna Mae Diehl1, Michael A Morse14 and Gayathri R Devi34*

Author affiliations

1 Department of Medicine, 2606 Duke University Medical Center, Durham, NC, 27710, USA

2 Biomanufacturing Research Institute and Technology Enterprise (BRITE), Department of Pharmaceutical Sciences, Durham, North Carolina Central University, Durham, NC, 27707, USA

3 Department of Surgery, Duke University Medical Center, Durham, NC, 27710, USA

4 Duke Cancer Institute, Duke University Medical Center, Durham, NC, 27710, USA

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Citation and License

BMC Cancer 2012, 12:402  doi:10.1186/1471-2407-12-402

Published: 10 September 2012

Abstract

Background

Crosstalk between malignant hepatocytes and the surrounding peritumoral stroma is a key modulator of hepatocarcinogenesis and therapeutic resistance. To examine the chemotherapy resistance of these two cellular compartments in vitro, we evaluated a well-established hepatic tumor cell line, HepG2, and an adult hepatic stellate cell line, LX2. The aim was to compare the chemosensitization potential of arsenic trioxide (ATO) in combination with sorafenib or fluorouracil (5-FU), in both hepatic tumor cells and stromal cells.

Methods

Cytotoxicity of ATO, 5-FU, and sorafenib, alone and in combination against HepG2 cells and LX2 cells was measured by an automated high throughput cell-based proliferation assay. Changes in survival and apoptotic signaling pathways were analyzed by flow cytometry and western blot. Gene expression of the 5-FU metabolic enzyme, thymidylate synthase, was analyzed by real time PCR.

Results

Both HepG2 and LX2 cell lines were susceptible to single agent sorafenib and ATO at 24 hr (ATO IC50: 5.3 μM in LX2; 32.7 μM in HepG2; Sorafenib IC50: 11.8 μM in LX2; 9.9 μM in HepG2). In contrast, 5-FU cytotoxicity required higher concentrations and prolonged (48–72 hr) drug exposure. Concurrent ATO and 5-FU treatment of HepG2 cells was synergistic, leading to increased cytotoxicity due in part to modulation of thymidylate synthase levels by ATO. Concurrent ATO and sorafenib treatment showed a trend towards increased HepG2 cytotoxicity, possibly due to a significant decrease in MAPK activation in comparison to treatment with ATO alone.

Conclusions

ATO differentially sensitizes hepatic tumor cells and adult hepatic stellate cells to 5-FU and sorafenib. Given the importance of both of these cell types in hepatocarcinogenesis, these data have implications for the rational development of anti-cancer therapy combinations for the treatment of hepatocellular carcinoma (HCC).

Keywords:
Hepatocellular carcinoma; Arsenic trioxide; Sorafenib; 5-fluorouracil; Apoptosis; High throughput assay; Stroma