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

Author(s): Jones RJ, Boyce T, Fennell M, Jacobs V, Pinto F, Duffield E, Clack G, Green T, Kelly J, Robertson J

Publication: Cancer Chemother Pharmacol, 2008, Vol. 61, Page 23-32

PubMed ID: 17909809 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine if prolonged cold ischemia of breast biopsy specimens altered levels of the following established Src kinase targets: phospho-Paxillin, phospho-FAK and total phospho-Tyrosine, as well as the ratio of phospho-Paxillin to total Paxillin. Specimens were stored at room temperature on damp blotting paper prior to cryopreservation by immersion on liquid nitrogen. Although breast biopsy specimens were collected from a total of 20 patients, the sample size for each timepoint ranged from 3 to 10 patients.

   Summary of Findings:

   Significant increases in the levels of phospho-Paxillin (100%) and total phospho-Tyrosine (350%), as well as the ratio of phospho-Paxillin to total Paxillin were observed in breast biopsy specimens after a cold ischemia time of 60 min compared to 0 min controls. Minute fluctuations in total Paxillin, and phospho-FAK levels after cold ischemia of 60 min were nonsignificant. Cold ischemia of less than 60 min did not affect any of the phospho-proteins analyzed.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunoassay

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Cold ischemia time	0 min 10 min 20 min 30 min 45 min 60 min
   Immunoassay Specific	Targeted peptide/protein	Paxillin Phospho-Paxillin Phospho-FAK Total phospho-Tyrosine

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2. Study Purpose

   The purpose of this study was to determine if prolonged cold ischemia of surgically resected bladder specimens altered levels of the following established Src kinase targets: phospho-Paxillin, phospho-FAK and total phospho-Tyrosine, as well as the ratio of phospho-Paxillin to total Paxillin. Specimens were stored at room temperature in normal saline prior to cryopreservation by immersion in liquid nitrogen. Although bladder specimens were collected from a total of 5 patients, the sample size for each timepoint ranged from 4 to 5 patients.

   Summary of Findings:

   Bladder specimens subjected to cold ischemia at room temperature in normal saline for up to 45 min did not exhibit any significant or robust changes in expression of phospho- or total Paxillin, phospho-FAK, or total phospho-Tyrosine.

   Biospecimens

   • Tissue - Bladder

   Preservative Types

   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunoassay

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Cold ischemia time	0 min 10 min 20 min 30 min 45 min
   Immunoassay Specific	Targeted peptide/protein	Paxillin phospho-Paxillin phospho-FAK Total phospho-Tyrosine

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