Quantitative assessment of effect of preanalytic cold ischemic time on protein expression in breast cancer tissues.
Author(s): Neumeister VM, Anagnostou V, Siddiqui S, England AM, Zarrella ER, Vassilakopoulou M, Parisi F, Kluger Y, Hicks DG, Rimm DL
Publication: J Natl Cancer Inst, 2012, Vol. 104, Page 1815-24
PubMed ID: 23090068 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of cold ischemia time on the antigenicity of 23 proteins among formalin-fixed paraffin-embedded (FFPE) breast cancer resection and biopsy specimens.
Conclusion of Paper
Of the 23 antigens examined on tissue microarrays, only histone 4 (HIST4H4) and phosphorylated tyrosine (4G10) immunostaining significantly decreased with longer cold ischemia times while acetylated lysine immunostaining significantly increased. Non-significant changes were observed for 11 antigens, and cold ischemia time had no effect on the remaining 9 antigens among the 93 resection specimens on the tissue microarray. Notably, the common breast cancer markers estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and Ki67 showed no significantly altered antigenicity with increasing cold ischemia time, although ER and PR showed decreasing trends among the tissue microarray cohort. The decrease in phosphorylated tyrosine (4G10) immunostaining with longer cold ischemia times was confirmed in matched biopsy and surgical resection specimens, but the decrease in HIST4H4 immunostaining was not observed, despite the difference in cold ischemia between resection and biopsy specimens. Interestingly, although only shown to have a trend towards increasing with longer cold ischemia times on the tissue microarray, HIF-1-alpha (HIF1A) and A kinase (PRKA) anchor protein 13 (AKAP13) had significantly increased immunostaining in the resection specimens (60-180 min cold ischemia) compared to the biopsies (<3 min cold ischemia).
Studies
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Study Purpose
The purpose of this study was to determine the effects of cold ischemia time on the antigenicity of 23 proteins among FFPE breast cancer resection and biopsy specimens. Two cohorts of specimens were examined, the first consisting of 93 resection specimens with recorded cold ischemia times in the range of 25-415 min on a tissue microarray, and the second consisting of whole slides for 25 matched pairs of surgical resections (estimated cold ischemia time of 60-180 min) and core needle biopsies (estimated cold ischemia time of <3 min). The automated quantitative analysis (AQUA) system was used to score immunofluorescence.
Summary of Findings:
Among the tissue microarray cohort of specimens, although trends towards changes in immunostaining with increased cold ischemia time were noted for 11/23 antigens, only HIST4H4 and phosphorylated tyrosine (4G10) immunostaining significantly decreased with longer cold ischemia times while acetylated lysine immunostaining significantly increased. Cold ischemia time had no effect on the remaining 9 antigens among the microarray specimens. When a subset of 9 antibodies (acetylated lysine not included) was used to evaluate the cohort of matched resection and biopsy specimens with the associated differences in cold ischemia time, phosphorylated tyrosine (4G10) immunostaining was lower among the surgical resection specimens than the biopsies, but HIST4H4 immunostaining showed no differences. Interestingly, although only shown to have a trend towards increasing with longer cold ischemia times on the tissue microarray, HIF1A and AKAP13 had significantly increased immunostaining among the resection specimens than the biopsies. Notably, the common breast cancer markers ER, PR, HER2, and Ki67 showed no significantly altered antigenicity with increasing cold ischemia time, although ER and PR showed decreasing trends among the tissue microarray cohort.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Protein Tissue microarray Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Cold ischemia time <3 min
60-180 min
25-415 min
Biospecimen Aliquots and Components Type of slide Tissue microarray
Whole section slide
Biospecimen Acquisition Method of tissue acquisition Biopsy
Surgical resection