Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision
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 a...
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| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2004-06-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/04366RR04 |
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| author | Annika Spruessel Garnet Steimann Mira Jung Sung A. Lee Theresa Carr Anne-Kristin Fentz Joerg Spangenberg Carsten Zornig Hartmut H. Juhl Kerstin A. David |
| author_facet | Annika Spruessel Garnet Steimann Mira Jung Sung A. Lee Theresa Carr Anne-Kristin Fentz Joerg Spangenberg Carsten Zornig Hartmut H. Juhl Kerstin A. David |
| author_sort | Annika Spruessel |
| collection | DOAJ |
| description | 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 IMAC3Ni 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. |
| format | Article |
| id | doaj-art-70f9eb53bba445498791654841dcac71 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2004-06-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-70f9eb53bba445498791654841dcac712025-08-20T02:26:07ZengTaylor & Francis GroupBioTechniques0736-62051940-98182004-06-013661030103710.2144/04366RR04Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excisionAnnika Spruessel0Garnet Steimann1Mira Jung2Sung A. Lee3Theresa Carr4Anne-Kristin Fentz5Joerg Spangenberg6Carsten Zornig7Hartmut H. Juhl8Kerstin A. David91Center for Cancer Research at the Israelitic Hospital, Hamburg, Germany1Center for Cancer Research at the Israelitic Hospital, Hamburg, Germany2Georgetown University, Washington, DC, USA2Georgetown University, Washington, DC, USA2Georgetown University, Washington, DC, USA1Center for Cancer Research at the Israelitic Hospital, Hamburg, Germany1Center for Cancer Research at the Israelitic Hospital, Hamburg, Germany3Israelitisches Krankenhaus, Hamburg, Germany1Center for Cancer Research at the Israelitic Hospital, Hamburg, Germany1Center for Cancer Research at the Israelitic Hospital, Hamburg, GermanyThe 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 IMAC3Ni 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.https://www.future-science.com/doi/10.2144/04366RR04 |
| spellingShingle | Annika Spruessel Garnet Steimann Mira Jung Sung A. Lee Theresa Carr Anne-Kristin Fentz Joerg Spangenberg Carsten Zornig Hartmut H. Juhl Kerstin A. David Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision BioTechniques |
| title | Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision |
| title_full | Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision |
| title_fullStr | Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision |
| title_full_unstemmed | Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision |
| title_short | Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision |
| title_sort | tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision |
| url | https://www.future-science.com/doi/10.2144/04366RR04 |
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