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Knockdown of ornithine decarboxylase antizyme 1 causes loss of uptake regulation leading to increased N 1, N 11-bis(ethyl)norspermine (BENSpm) accumulation and toxicity in NCI H157 lung cancer cells

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Abstract

Ornithine decarboxylase antizyme 1 (AZ1) is a major regulatory protein responsible for the regulation and degradation of ornithine decarboxylase (ODC). To better understand the role of AZ1 in polyamine metabolism and in modulating the response to anticancer polyamine analogues, a small interfering RNA strategy was used to create a series of stable clones in human H157 non-small cell lung cancer cells that expressed less than 5–10% of basal AZ1 levels. Antizyme 1 knockdown clones accumulated greater amounts of the polyamine analogue N 1,N 11-bis(ethyl)norspermine (BENSpm) and were more sensitive to analogue treatment. The possibility of a loss of polyamine uptake regulation in the knockdown clones was confirmed by polyamine uptake analysis. These results are consistent with the hypothesis that AZ1 knockdown leads to dysregulation of polyamine uptake, resulting in increased analogue accumulation and toxicity. Importantly, there appears to be little difference between AZ1 knockdown cells and cells with normal levels of AZ1 with respect to ODC regulation, suggesting that another regulatory protein, potentially AZ2, compensates for the loss of AZ1. The results of these studies are important for the understanding of both the regulation of polyamine homeostasis and in understanding the factors that regulate tumor cell sensitivity to the anti-tumor polyamine analogues.

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Abbreviations

AZ1:

Ornithine decarboxylase antizyme 1

ODC:

Ornithine decarboxylase

NSCLC:

Non-small cell lung cancer

BENSpm:

N 1,N 11-bis(ethyl)norspermine

AZin:

Ornithine decarboxylase antizyme inhibitor

siRNA:

Small interfering RNA

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Acknowledgments

Anti-ODC primary antibody was a kind gift from Dr. Lisa Shantz, Pennsylvania State University. These studies were supported by National Institutes of Health grants CA 51085, CA 98454, and CA 149095.

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  1. Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Bunting-Blaustein Cancer Research Building 1, Room 551, 1650 Orleans Street, Baltimore, MD, 21231, USA

    Alison V. Fraser, Andrew C. Goodwin, Amy Hacker-Prietz, Elizabeth Sugar & Robert A. Casero Jr.

  2. Department of Pharmaceutical and Biomedical Sciences, Medical University of South Carolina, 280 Calhoun St, Charleston, SC, 29425, USA

    Patrick M. Woster

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  1. Alison V. Fraser
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  2. Andrew C. Goodwin
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Correspondence to Robert A. Casero Jr..

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Fraser, A.V., Goodwin, A.C., Hacker-Prietz, A. et al. Knockdown of ornithine decarboxylase antizyme 1 causes loss of uptake regulation leading to increased N 1, N 11-bis(ethyl)norspermine (BENSpm) accumulation and toxicity in NCI H157 lung cancer cells. Amino Acids 42, 529–538 (2012). https://doi.org/10.1007/s00726-011-1030-z

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