Azacytidine sensitizes acute myeloid leukemia cells to arsenic trioxide by up-regulating the arsenic transporter aquaglyceroporin 9

Journal of Hematology & Oncology

Eric Tse¹  Yok-Lam Kwong¹  Sidney Tam³  Bonnie Fong³  Yuen-Yee Cheng²  Thomas Sau-Yan Chan¹  Karen Ng¹  David Chau¹ 

[1] Division of Haematology, Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pok Fu Lam, Hong Kong, Hong Kong;Asbestos Diseases Research Institute, ADRI Bernie Banton Centre, University of Sydney, Concord Hospital, Sydney, Australia;Department of Pathology and Clinical Biochemistry, Queen Mary Hospital, Pok Fu Lam, Hong Kong, Hong Kong

关键词: Aquaglyceroporin 9;    Demethylating agents;    Azacytidine;    Arsenic trioxide;    Acute myeloid leukemia;   

Others  :  1211896
DOI  :  10.1186/s13045-015-0143-3

 received in 2015-01-16, accepted in 2015-04-27,  发布年份 2015

【 摘 要 】

Background

The therapeutic efficacy of arsenic trioxide (As₂O₃) in acute myeloid leukemia (AML) is modest, which is partly related to its limited intracellular uptake into the leukemic cells. As₂O₃ enters cells via the transmembrane protein aquaglyceroporin 9 (AQP9). Azacytidine, a demethylating agent that is approved for the treatment of AML, has been shown to have synergistic effect with As₂O₃. We tested the hypothesis that azacytidine might up-regulate AQP9 and enhances As₂O₃-mediated cytotoxicity in AML.

Methods

Arsenic-induced cytotoxicity, the expression of AQP9, and the intracellular uptake of As₂O₃ were determined in AML cell lines and primary AML cells with or without azacytidine pre-treatment. The mechanism of AQP9 up-regulation was then investigated by examining the expression of transcription factors for AQP9 gene and the methylation status of their gene promoters.

Results

As₂O₃-induced cytotoxicity in AML cell lines was significantly enhanced after azacytidine pre-treatment as a result of AQP9 up-regulation, leading to increased arsenic uptake and hence intracellular concentration. Blocking AQP9-mediated As₂O₃ uptake with mercury chloride abrogated the sensitization effect of azacytidine. AQP9 promoter does not contain CpG islands. Instead, azacytidine pre-treatment led to increased expression of HNF1A, a transcription activator of AQP9, through demethylation of HNF1A promoter. HNF1 knockdown abrogated azacytidine-induced AQP9 up-regulation and almost completely blocked intracellular As₂O₃ entry, confirming that azacytidine enhanced As₂O₃-mediated cell death via up-regulation of HNF1A and hence increased AQP9 and As₂O₃ intracellular concentration. Azacytidine sensitization to As₂O₃ treatment was re-capitulated also in primary AML samples. Finally, azacytidine did not enhance arsenic toxicity in a liver cell line, where HNF1A was largely unmethylated.

Conclusions

Azacytidine sensitizes AML cells to As₂O₃ treatment, and our results provide proof-of-principle evidence that pharmacological up-regulation of AQP9 potentially expands the therapeutic spectrum of As₂O₃. Further clinical trial should evaluate the efficacy of azacytidine in combination with As₂O₃ in the treatment of AML.

【 授权许可】

   
2015 Chau et al.; licensee BioMed Central.

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