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J Histochem Cytochem ; 67(3): 159-168, 2019 03.
Article in English | MEDLINE | ID: mdl-30562131

ABSTRACT

Although there are thousands of formalin-fixed paraffin-embedded (FFPE) tissue blocks potentially available for scientific research, many are of questionable quality, partly due to unknown preanalytical variables. We analyzed FFPE tissue biospecimens as part of the National Cancer Institute (NCI) Biospecimen Preanalytical Variables program to identify mRNA markers denoting cold ischemic time. The mRNA was extracted from colon, kidney, and ovary cancer FFPE blocks (40 patients, 10-12 hr fixation time) with 1, 2, 3, and 12 hr cold ischemic times, then analyzed using qRT-PCR for 23 genes selected following a literature search. No genes tested could determine short ischemic times (1-3 hr). However, a combination of three unstable genes normalized to a more stable gene could generate a "Cold Ischemia Score" that could distinguish 1 to 3 hr cold ischemia from 12 hr cold ischemia with 62% sensitivity and 84% specificity.


Subject(s)
Cold Ischemia/methods , Colonic Neoplasms/genetics , Kidney Neoplasms/genetics , Neoplasm Proteins/genetics , Ovarian Neoplasms/genetics , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , Female , Fixatives/chemistry , Formaldehyde/chemistry , Humans , Kidney Neoplasms/metabolism , Kidney Neoplasms/pathology , Male , Neoplasm Proteins/metabolism , Ovarian Neoplasms/metabolism , Ovarian Neoplasms/pathology , Paraffin Embedding/methods , RNA, Messenger/metabolism , Time Factors , Tissue Fixation/methods , Transcriptome
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