The influence of tissue ischemia on biomarker expression in colorectal cancer.
Author(s): Havelund BM, Olsen DA, Andersen RF, Spindler KL, Brandslund I, Jakobsen A, Soerensen FB
Publication: Appl Immunohistochem Mol Morphol, 2013, Vol. 21, Page 298-307
PubMed ID: 23060299 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of ischemia time on HIF-1alpha mRNA expression in colorectal cancer tissue collected from 25 patients. All specimens were exposed to perioperative ischemia, defined by the authors as the time between ligation of blood flow to the tumor and receipt by the pathology department. Specimens were then either immediately placed in RNAlater or subjected to an additional period of cold ischemia in a room temperature air tight container with physiological saline. Specimens were stored in RNAlater at 4 degrees C for 24 hours before being transferred to -20 degrees C until analysis.
Summary of Findings:
A cold ischemia time of 30 or 60 min, which was in addition to the recorded perioperative ischemia time, did not have a significant effect on HIF-1alpha mRNA levels compared to case-matched specimens that were immediately preserved upon arrival in the pathology department. Furthermore, no correlation was found between perioperative or total ischemia time and HIF-1alpha gene expression.
Biospecimens
Preservative Types
- RNAlater
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform RNA Real-time qRT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Cold ischemia time 0 min
30 min
60 min
Biospecimen Acquisition Unspecified ischemia time 22-240 min
Real-time qRT-PCR Specific Targeted nucleic acid HIF-1alpha
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Study Purpose
The purpose of this study was to determine the effects of ischemia time on HIF-1alpha protein concentration in colorectal cancer tissue collected from 25 patients. All specimens were exposed to perioperative ischemia, defined by the authors as the time between ligation of blood flow to the tumor and receipt by the pathology department. Specimens were then either immediately snap frozen in liquid nitrogen or subjected to an additional period of cold ischemia in a room temperature air tight container with physiological saline. Specimens were stored at -80 degrees C until analysis.
Summary of Findings:
A cold ischemia time of 30 min, but not 60 min, significantly increased HIF-1alpha protein concentration compared to specimens that were immediately snap-frozen upon receipt and dissection in the pathology lab (p=0.03). Elimination of one potential outlier tumor reduced the statistical strength of the change observed at 30 min cold ischemia (p=0.047). There was no relationship between HIF-1alpha protein concentration and the duration of perioperative or total ischemia.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Protein Colorimetric assay Protein ELISA Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Cold ischemia time 0 min
30 min
60 min
Biospecimen Acquisition Unspecified ischemia time 22-240 min
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Study Purpose
The purpose of this study was to determine the effects of cold and total ischemia time on HIF-1alpha, GLUT-1, Bcl-2, and Ki-67 IHC staining in FFPE colorectal cancer tissue from 25 patients. Potential differences in IRS between preoperative biopsies and whole sections were also examined. With the exception of preoperative biopsies, all specimens were exposed to perioperative ischemia, defined by the authors as the time between ligation of blood flow to the tumor and receipt by the pathology department. Specimens were then either immediately immersed in formalin or subjected to an additional period of cold ischemia in a room temperature air tight container with physiological saline.
Summary of Findings:
Overall, 30 or 60 minutes of cold ischemia did not significantly affect HIF-1alpha, GLUT-1, or Ki-67 IRS, or the intensity of Bcl-2 immunostaining, however a large degree of intra-individual variation was observed for six specimens. For these six specimens, HIF-1alpha and GLUT-1 IRS ranged between 2 and 5 for the pre-operative biopsy, the whole section, and the three different cold ischemia time points for the same specimen. A relationship between IRS and perioperative or total ischemia time was not observed for any antigen investigated. IRS did not differ between preoperative biopsies and whole sections for any of the antigens investigated, although the authors report that HIF-1alpha, GLUT-1, and Ki-67 IRS tended to be lower in whole sections when compared to biopsies.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Cold ischemia time 0 min
30 min
60 min
Immunohistochemistry Specific Targeted peptide/protein HIF-1alpha
GLUT-1
Ki-67
Bcl-2
Biospecimen Acquisition Unspecified ischemia time 22-240 min
Biospecimen Acquisition Method of tissue acquisition Biopsy
Surgical resection