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Aneuploid human colonic epithelial cells are sensitive to AICAR-induced growth inhibition through EGFR degradation

Oncogene volume 32, pages 3139–3146 (2013)Cite this article

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Abstract

Trisomy for chromosome 7 is frequently observed as an initiating event in sporadic colorectal cancer. Although unstable chromosome numbers and recurrent aneuploidies drive a large fraction of human cancers, targeted therapies selective to pre-neoplastic trisomic cells are non-existent. We have previously characterized a trisomy 7 cell line (1CT+7) spontaneously derived from normal diploid human colonic epithelial cells that aberrantly expresses the epidermal growth factor receptor (EGFR, chromosome 7p11). Recent studies identified AICAR (5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside) as a pharmacological inhibitor of aneuploid murine fibroblast proliferation. Here, we report that AICAR induces profound cytostatic and metabolic effects on 1CT+7 cells, but not on their isogenic diploid counterpart. Dose–response experiments indicate that 1CT+7 cells are fourfold preferentially sensitive to AICAR compared to diploid cells. Unexpectedly, treatment of 1CT+7 cells with AICAR led to a reversible 3.5-fold reduction (P=0.0025) in EGFR overexpression. AICAR-induced depletion of EGFR protein can be abrogated through inhibition of the proteasome with MG132. AICAR also heavily promoted EGFR ubiquitination in cell-based immunoprecipitation assays, suggesting enhanced degradation of EGFR protein mediated by the proteasome. Moreover, treatment with AICAR reduced EGFR protein levels in a panel of human colorectal cancer cells in vitro and in xenograft tumors in vivo. Our data collectively support the pharmacological compound AICAR as a novel inhibitor of EGFR protein abundance and as a potential anticancer agent for aneuploidy-driven colorectal cancer.

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Acknowledgements

We thank RY Quach for figure artwork and U Eskiocak, G Stadler and AR Mullen (UT Southwestern) for technical assistance. We also express gratitude to YC Tang and A Amon (MIT) for providing xenograft tumors for IHC analysis and for critical reading of the manuscript. HA-tagged ubiquitin plasmids and anti-HA antibodies were a kind gift from LJ Huang (UT Southwestern). EGFR-V5 expression constructs and human bronchial epithelial cell-EGFR cell lines were provided by C Nirodi (UT Southwestern). This work was supported by CPRIT Training Grant RP101496 to PL, NASA Grants NNX09AU95G, NNX11AC15G, and NNX11AC54G and NCI SPORE CA70907 to JWS.

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Authors and Affiliations

  1. Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA

    P Ly, S B Kim, A A Kaisani, G Marian, W E Wright & J W Shay

  2. Center of Excellence in Genomic Medicine Research, King Abdulaziz University, Jeddah, Saudi Arabia

    J W Shay

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  1. P Ly
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Correspondence to J W Shay.

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Ly, P., Kim, S., Kaisani, A. et al. Aneuploid human colonic epithelial cells are sensitive to AICAR-induced growth inhibition through EGFR degradation. Oncogene 32, 3139–3146 (2013). https://doi.org/10.1038/onc.2012.339

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