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The impact of delay in cryo-fixation on biomarkers of Src tyrosine kinase activity in human breast and bladder cancers

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Abstract

Demonstration of pharmacodynamic activity of new, targeted cancer drugs in tumour tissue is potentially important in guiding early drug development. However, delays between tumour sampling and sample fixation may result in variability of pharmacodynamic biomarkers. The aim of this study, was to assess the impact of delays in fixation on biomarkers of Src kinase activity. A total of 20 patients with locally advanced breast cancer and 5 with early bladder cancer had multiple tissue samples taken which were fixed at documented time points up to 60 min after biopsy. These were examined to determine if the amount of Paxillin, phospho-Paxillin, phospho-focal adhesion kinase (FAK) and total phospho-Tyrosine changed over time, using a quantitative lysate immunoassay. In breast cancer, there was an increase in the amount of phospho-Paxillin (60% per h; P = 0.019) up to 60 min after biopsy. The amount of total Paxillin decreased (28% per h; P = 0.034) over the same time course. In early bladder cancer, no changes were noted in any endpoints up to 45 min. Standardisation of the time taken between biopsy and fixation may be critical, particularly in studies using phosphorylated protein biomarkers.

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Acknowledgements

Pamela de Clive Lowe (research nurse), Karina Meachin (study manager).

Competing interests

MF, VJ, ED, TG and GC are employees of AstraZeneca plc. RJ is a consultant to AstraZeneca plc.

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Authors and Affiliations

  1. Beatson Oncology Centre, Western Infirmary of Glasgow, Glasgow, G11 6NT, UK

    Rob J. Jones

  2. AstraZeneca, Macclesfield, UK

    Michael Fennell, Vivien Jacobs, Emma Duffield, Glen Clack & Tim Green

  3. City Hospital, Nottingham, UK

    Tamsin Boyce & John Robertson

  4. Addenbrooke’s Hospital, Cambridge, UK

    Francesco Pinto & John Kelly

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  1. Rob J. Jones
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  2. Tamsin Boyce
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  3. Michael Fennell
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  4. Vivien Jacobs
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  5. Francesco Pinto
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Correspondence to Rob J. Jones.

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The work presented was supported by AstraZeneca plc.

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Jones, R.J., Boyce, T., Fennell, M. et al. The impact of delay in cryo-fixation on biomarkers of Src tyrosine kinase activity in human breast and bladder cancers. Cancer Chemother Pharmacol 61, 23–32 (2008). https://doi.org/10.1007/s00280-007-0440-9

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