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Arsenic trioxide inhibits the growth of Adriamycin resistant osteosarcoma cells through inducing apoptosis

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Abstract

Given that arsenic trioxide (As2O3) has been successfully used as a chemotherapeutic agent for refractory malignant tumors, this study is aimed at investigating the effect of As2O3 on human Adriamycin resistant osteosarcoma cell line Saos-2. The mechanism underlying multi drug resistance (MDR) in osteosarcoma cells and the anti-tumor effect of As2O3 on Adriamycin resistant osteosarcoma cells were analyzed. In our experiment, we first selected Adriamycin resistant osteosarcoma cell line by growing the classic osteosarcoma cell line Saos-2 in the medium with increasing drug concentrations. Then, we compared the IC50s of the osteosarcoma cells treated with different anticancer drugs by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Subsequently, we assessed the expression of classic MDR related molecules, Pgp, multidrug resistance-associated protein (MRP) and glutathione (GSH) activity in the wild type and Adriamycin resistant Saos-2 cells. Furthermore, the apoptosis was assessed by concerning DNA fragment and flow cytometry with Annexin-V staining. To elucidate the underlying mechanism of the apoptosis, related proteins Bcl-2, Bcl-xL, Bax, Bak, cleaved Caspase-3 and cleaved Caspase-9 were analyzed by western blotting. The data showed that the resistance to Adriamycin affected the sensitivity of osteosarcoma cell to other chemotherapeutic agents. The IC50s of Saos-2/ADM cells for methotrexate (1.74-fold), Cisplatin (1.43-fold) and As2O3 (1.21-fold) were increased compared with Saos-2 control cells. The expression of Pgp was upregulated comparing with the control cells. No significant difference was detected about the MRP and the glutathione-S-transferase activity and intracellular GSH concentration among different treated osteosarcoma cells. Apoptosis was observed and proved. The western blotting showed that the expression of Bcl-2 and Bcl-xL was downregulated. Meanwhile, the level of Bax, Bak, cleaved Caspase-3 and cleaved Caspase-9 was upregulated after treated with As2O3. The study suggests that Adriamycin resistant osteosarcoma cells have good response to As2O3-based chemotherapy in vitro, probably via the pathway of inducing apoptosis. And As2O3 might serve as an excellent alternative candidate for adjuvant chemotherapeutic agent on this incurable pediatric sarcoma.

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Acknowledgments

One or more of the authors (Guo Wei, MD, PhD) have received funding from the Capital Development Foundation of Beijing (Grant no. 2005-1009).

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Authors and Affiliations

  1. Musculoskeletal Tumor Center, Peking University People’s Hospital, #11 South Xizhimen Street, 100044, Beijing, People’s Republic of China

    Hui Zhao, Wei Guo, Changliang Peng, Tao Ji & Xinchang Lu

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  1. Hui Zhao
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  2. Wei Guo
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  3. Changliang Peng
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  4. Tao Ji
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  5. Xinchang Lu
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Correspondence to Wei Guo.

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Each author certifies that his or her institution has approved the experimental protocol for this investigation and that all investigations were conducted in conformity with ethical principles of research.

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Zhao, H., Guo, W., Peng, C. et al. Arsenic trioxide inhibits the growth of Adriamycin resistant osteosarcoma cells through inducing apoptosis. Mol Biol Rep 37, 2509–2515 (2010). https://doi.org/10.1007/s11033-009-9765-2

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  • DOI: https://doi.org/10.1007/s11033-009-9765-2

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