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Downregulation of the type 1 insulin-like growth factor receptor in mouse melanoma cells is associated with enhanced radiosensitivity and impaired activation of Atm kinase

Oncogene volume 20, pages 4029–4040 (2001)Cite this article

Abstract

The type 1 insulin-like growth factor receptor (IGF1R) is required for growth, tumorigenicity and protection from apoptosis. IGF1R overexpression is associated with radioresistance in breast cancer. We used antisense (AS) RNA to downregulate IGF1R expression in mouse melanoma cells. Cells expressing AS-IGF1R transcripts were more radiosensitive in vitro and in vivo than controls. Also they showed reduced radiation-induced p53 accumulation and p53 serine 18 phosphorylation, and radioresistant DNA synthesis. These changes were reminiscent of the cellular phenotype of the human genetic disorder ataxia-telangiectasia (A-T), caused by mutations in the ATM gene. Cellular Atm protein levels were lower in AS-IGF1R-transfected cells than in control cells, although there was no difference in Atm expression at the transcriptional level. AS-IGF1R cells had detectable basal Atm kinase activity, but failed to induce kinase activity after irradiation. This suggests that IGF1R signalling can modulate the function of Atm, and supports the concept of targeted IGF1R downregulation as a potential treatment for malignant melanoma and other radioresistant tumours.

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Abbreviations

A-T:

ataxia telangiectasia

ATM:

ataxia-telangiectasia mutated

DMEM:

Dulbecco's modification of Eagle's medium

DTT:

dithiothreitol

EGF:

epidermal growth factor

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

IGF:

insulin-like growth factor

IGF1R:

type 1 IGF receptor

IR:

irradiation

MEFs:

mouse embryo fibroblasts

PAGE:

polyacrylamide gel electrophoresis

PBMCs:

peripheral blood mononuclear cells

PBSA:

phosphate buffered saline

PCR:

polymerase chain reaction

PHA:

phytohaemagglutin

PMSF:

phenylmethylsulphonyl fluoride

RDS:

radio-resistant DNA synthesis.

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Acknowledgements

We are grateful to Ian Hart for B16.F1 cells, Renato Baserga for the HspA plasmid, Philip Leder for ap29 and ap24 cells, Sandra Peake for technical assistance and Sally Davies for p53 1–66 protein. We are also grateful to Ian Hickson, Chris Norbury, Adrian Harris, Penny Jeggo and Heather Beamish for advice, and Haim Werner for comments on the manuscript. This study was supported by the ICRF and by MRC Clinician Scientist Fellowship G108/191. Work in the laboratory of Y Shiloh was supported by the A-T Children's Project, The A-T Medical Research Foundation and NIH grant R01 NS31763.

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Authors and Affiliations

  1. IGF Group, Molecular Oncology Laboratories, Weatherall Institute of Molecular Medicine, Oxford, OX3 9DS, UK

    V M Macaulay, A J Salisbury, E A Bohula & M P Playford

  2. Hybridoma Unit, The George S. Wise Faculty of Life Science, Tel Aviv University, Tel Aviv, 69978, Israel

    N I Smorodinsky

  3. Department of Human Genetics and Molecular Medicine, Sackler School of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel

    Y Shiloh

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  1. V M Macaulay
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Correspondence to V M Macaulay.

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Macaulay, V., Salisbury, A., Bohula, E. et al. Downregulation of the type 1 insulin-like growth factor receptor in mouse melanoma cells is associated with enhanced radiosensitivity and impaired activation of Atm kinase. Oncogene 20, 4029–4040 (2001). https://doi.org/10.1038/sj.onc.1204565

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