Open AccessTissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision
Author(s) -
Annika Spruessel,
Garnet Steimann,
Mira Jung,
Sung A Lee,
Theresa D. Carr,
Anne-Kristin Fentz,
Jörg Spangenberg,
C. Zornig,
Hartmut Juhl,
Kerstin A. David
Publication year - 2004
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/04366rr04
Subject(s) - gene expression , gene , microbiology and biotechnology , real time polymerase chain reaction , ischemia , messenger rna , gene expression profiling , reverse transcription polymerase chain reaction , microarray , pathology , dna microarray , biology , medicine , genetics
The aim of this study was to determine the impact of ischemia on gene and protein expression profiles of healthy and malignant colon tissue and, thus, on screening studies for identification of molecular targets and diagnostic molecular patterns. Healthy and malignant colon tissue were snap-frozen at various time points (3–30 min) after colon resection. Gene and protein expression were determined by microarray (HG-U133A chips) and surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) technology (CM10 chips, SAX2 chips, and IMAC3 Ni chips), respectively. Real-time reverse transcription PCR (RT-PCR) was used for comparative measurement of expression of particular genes. Initial changes of gene and protein expression profiles were already observed 5–8 min after colon resection. Fifteen minutes after surgery, 10%–15% of molecules, and after 30 min, 20% of all detectable genes and proteins, respectively, differed significantly from the baseline values. Significant changes of expression were found in most functional groups. As confirmed by real-time RT-PCR, this included not only known hypoxia-related molecules (HIF-1α, c-fos, HO-1) but also cytoskeletal genes (e.g., CK20) and tumor-associated antigens (e.g., CEA). In conclusion, preanalytical factors, such as tissue ischemia time, dramatically affect molecular data. Control of these variables is mandatory to obtain reliable data in screening programs for molecular targets and diagnostic molecular patterns.