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Array comparative genomic hybridisation analysis of gamma-irradiated human thyrocytes

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Abstract

The susceptibility of thyroid epithelium to radiation-induced carcinogenesis is well recognised. In this context, thyroid carcinogenesis is associated with specific somatic ret/papillary thyroid carcinoma (PTC) rearrangements and morphologically with the papillary phenotype. Previous studies have demonstrated the possibility of inducing ret rearrangements in vitro using X-rays. The purpose of our study was to assess whether gamma (γ) radiation using a Caesium 137 source can induce specific ret rearrangements in a human thyroid epithelial cell culture model. We further hypothesised that if radiation-induced thyroid carcinogenesis is associated with non-random rearrangement events, then DNA copy gain and loss induced by irradiation may also occur in a non-random manner. We irradiated SV40-immortalised human thyroid epithelial cells with incremental doses of γ-radiation and, using TaqMan reverse-transcription polymerase chain reaction, looked for the presence of the common ret rearrangements. Cohorts showing evidence of ret/PTC chimeric transcripts were further analysed using microarray comparative genomic hybridisation (CGH) to detect copy gain and loss associated with radiation. Four Grays of γ-radiation was sufficient to induce ret/PTC-3. In this model, transcripts of ret/PTC-1 were not detected, and we suggest that the type of radiation may influence the resulting rearrangement that occurs. Using array CGH, we have demonstrated a predominant pattern of subtelomeric deletions occurring in association with this radiation cohort and raise the possibility that chromosome 10 may be a hotspot for radiation-induced damage for as yet unknown reasons.

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Acknowledgements

This work was supported by a grant from the Health Research Board of Ireland. Thanks to Professor Sissy Jhiang for providing ret/PTC-3 plasmid used for positive control. Thanks to Professor Kingston Mills for providing irradiation facilities.

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Authors and Affiliations

  1. Department of Histopathology Research, University of Dublin, Trinity College, Room 35/72, Institute of Molecular Medicine, Trinity Centre for Health Sciences, St. James’s Hospital , James’s Street, Dublin 8, Ireland

    Stephen P. Finn, Paul Smyth, Susanne Cahill, Richard Flavin, John O’Leary & Orla Sheils

  2. Department of Oral Pathology, Dublin Dental School and Hospital, Ireland

    Esther O’Regan

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  1. Stephen P. Finn
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  2. Paul Smyth
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  3. Esther O’Regan
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  4. Susanne Cahill
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  5. Richard Flavin
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  7. Orla Sheils
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Correspondence to Stephen P. Finn.

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Finn, S.P., Smyth, P., O’Regan, E. et al. Array comparative genomic hybridisation analysis of gamma-irradiated human thyrocytes. Virchows Arch 445, 396–404 (2004). https://doi.org/10.1007/s00428-004-1070-9

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  • DOI: https://doi.org/10.1007/s00428-004-1070-9

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