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Differential gene expression in human hepatocyte cell lines exposed to the antiretroviral agent zidovudine

  • Molecular Toxicology
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Abstract

Zidovudine (3′-azido-3′-deoxythymidine; AZT) is the most widely used nucleoside reverse transcriptase inhibitor for the treatment of AIDS patients and prevention of mother-to-child transmission of HIV-1. Previously, we demonstrated that AZT had significantly greater growth inhibitory effects upon the human liver carcinoma cell line HepG2 as compared to the immortalized human liver cell line THLE2. We have now used gene expression profiling to determine the molecular pathways associated with toxicity in both cell lines. HepG2 cells were incubated with 0, 2, 20, or 100 μM AZT for 2 weeks; THLE2 cells were treated with 0, 50, 500, or 2,500 μM AZT, concentrations that were equi-toxic to those used in the HepG2 cells. After the treatment, total RNA was isolated and subjected to microarray analysis. Global analysis of gene expression, with a false discovery rate ≤0.01 and a fold change ≥1.5, indicated that 6- to 70-fold more genes were differentially expressed in a significant concentration-dependent manner in HepG2 cells when compared to THLE2 cells. Comparative analysis indicated that 7 % of these genes were common to both cell lines. Among the common differentially expressed genes, 70 % changed in the same direction, most of which were associated with cell death and survival, cell cycle, cell growth and proliferation, and DNA replication, recombination, and repair. As determined by the uptake of [methyl-3H]AZT, the intracellular levels of total AZT were approximately twofold higher in THLE2 cells than in HepG2 cells. The expression of thymidine kinase 1 (TK1) and UDP-glucuronosyltransferase 2B7 (UGT2B7) genes that regulate the metabolic activation and deactivation of AZT, respectively, was increased in HepG2 cells but decreased in THLE2 cells after treatment with AZT. This differential response in AZT metabolism was confirmed by real-time PCR, western blotting, and/or enzymatic assays. These data indicate that molecular pathways involved with cell death and survival, cell cycle, cell growth and proliferation, and DNA replication, recombination, and repair are involved in the toxicities associated with AZT in both human cell lines, and that the difference in expression of TK1 and UGT2B7 in response to AZT treatment in HepG2 cells and THLE2 cells might explain why HepG2 cells are more sensitive than THLE2 cells to the toxicity of AZT.

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Acknowledgments

Qiangen Wu was supported by an appointment to the Postgraduate Research Program in the Division of Biochemical Toxicology at the National Center for Toxicological Research administered by Oak Ridge Institute for Science Education through an interagency agreement between the US Department of Energy and the FDA. This research was supported through an interagency agreement between the National Center for Toxicological Research, US Food and Drug Administration and the National Toxicology Program, National Institute of Environmental Health Sciences. (FDA IAG: 224-07-0007; NIH Y1ES1027).

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The authors declare that there are no conflicts of interest.

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  1. Qiangen Wu

    Present address: Department of Environmental Health, Indiana University, Bloomington, IN, 47405, USA

Authors and Affiliations

  1. Division of Biochemical Toxicology, National Center for Toxicological Research, Jefferson, AR, 72079, USA

    Jia-Long Fang, Qiangen Wu, Frederick A. Beland & Lei Guo

  2. Division of Systems Biology, National Center for Toxicological Research, Jefferson, AR, 72079, USA

    Tao Han & James C. Fuscoe

  3. Division of Bioinformatics and Biostatistics, National Center for Toxicological Research, Jefferson, AR, 72079, USA

    Ching-Wei Chang

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  1. Jia-Long Fang
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Correspondence to Jia-Long Fang.

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Fang, JL., Han, T., Wu, Q. et al. Differential gene expression in human hepatocyte cell lines exposed to the antiretroviral agent zidovudine. Arch Toxicol 88, 609–623 (2014). https://doi.org/10.1007/s00204-013-1169-3

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