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Effects of long-term storage on the detection of proteins, DNA, and mRNA in tissue microarray slides
Örebro University, School of Health and Medical Sciences.
Örebro University, School of Health and Medical Sciences, Örebro University, Sweden. Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.ORCID iD: 0000-0001-6881-237X
2011 (English)In: Journal of Histochemistry and Cytochemistry, ISSN 0022-1554, E-ISSN 1551-5044, Vol. 59, no 12, p. 1113-1121Article in journal (Refereed) Published
Abstract [en]

Storage of tissue slides has been claimed to induce dramatically reduced antigen detection particularly for immunohistochemistry (IHC). With tissue microarrays, the necessity to serially cut blocks in order to obtain as much material as possible is obvious. The presumed adverse effect of storage might hamper such an approach. The authors designed an experimental setting consisting of four different storage conditions with storage time of tissue slides of up to 1 year. Detection of proteins, DNA, and mRNA was performed using IHC and in situ hybridization techniques. Slight but significant changes in IHC occurred over time. The most important factor is the primary antibody used: four showed no significant changes, whereas limited decreases in 8 antibodies could be detected by image analysis. Whether the antigen was nuclear or cytoplasmic/membranous did not matter. No major differences between different storage conditions could be shown, but storage at 4C was overall the best procedure. Furthermore, gene copy number aberrations, chromosomal translocations, and the presence of mRNA could be detected on slides stored up to 1 year. In conclusion, in tissues optimally formalin fixed and using modern histological techniques, only minute changes in tissue antigenicity are induced by long-term storage.

Place, publisher, year, edition, pages
Sage Publications, 2011. Vol. 59, no 12, p. 1113-1121
Keywords [en]
immunohistochemistry, heat-induced antigen retrieval, antigenicity, formalin-fixed paraffin-embedded, FISH, CISH, tissue microarray
National Category
Medical Biotechnology
Research subject
Medicine
Identifiers
URN: urn:nbn:se:oru:diva-20595DOI: 10.1369/0022155411423779ISI: 000297649800006PubMedID: 22147608Scopus ID: 2-s2.0-84856012999OAI: oai:DiVA.org:oru-20595DiVA, id: diva2:467316
Available from: 2011-12-19 Created: 2011-12-19 Last updated: 2018-05-03Bibliographically approved
In thesis
1. Biomarkers in non-small cell lung carcinoma: methodological aspects and influence of gender, histology and smoking habits on estrogen receptor and epidermal growth factor family receptor signalling
Open this publication in new window or tab >>Biomarkers in non-small cell lung carcinoma: methodological aspects and influence of gender, histology and smoking habits on estrogen receptor and epidermal growth factor family receptor signalling
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Non-small cell lung carcinoma is a leading cause of cancer mortality worldwide. There are gender and smoking associated differences both in tumour types and clinical outcome. Squamous cell carcinomas (SCC) are more frequent among smoking men while females develop adenocarcinomas (ADCA). NSCLC among never smokers are mainly ADCA, and occurs mostly in females. The present thesis elucidates the role of estrogen receptor (ER) and epidermal growth factor receptor family (EGFR/HER2-4) in NSCLC in the perspective of gender and histology as well as the influence of smoking on those biomarkers.

A recently developed technique, tissue micro array (TMA), was employed.The question of how much of a tumour tissue that needed to be included in a TMA for biomarker analysis was analyzed by a statistical approach. Data indicates a sample size of three cylinders of tumour tissue with a diameter of 0.6 mm each as being appropriate and cost-effective. In order to optimally use the up to thousands of different tumour samples within a TMA, it would be optimal to serially cut and store slides before performing in situ detection of proteins and nucleic acids. Applying up to date methodology, and by evaluation with image analysis, data are presented that shows that such handling of TMA slides would be possible without any loss of biomarker information.

ERα is more frequently observed in ADCA and in females and a local estradiol synthesis is supported by the presence of aromatase. ERβ is identified as a positive prognostic marker in ADCA. Smoking is associated to increased levels of ERβ mRNA. EGFR over expression is associated with a ligand. Independent phosporylation of ERα. HER-4 intracellular domain may also act as a co-activator to ERα in ADCA, especially among neversmokers. The question of ER and EGFR family signalling crosstalk as a potential target for combined targeted therapy is raised.

Place, publisher, year, edition, pages
Örebro: Örebro universitet, 2011. p. 86
Series
Örebro Studies in Medicine, ISSN 1652-4063 ; 61
Keywords
Non-small cell lung carcinoma, estrogen receptor, epidermal growth factor receptor, HER-4, tissue microarray, immunohistochemistry, smoking habits, in situ hybridisation
National Category
Medical and Health Sciences Cancer and Oncology
Research subject
Oncology
Identifiers
urn:nbn:se:oru:diva-19725 (URN)978-91-7668-827-4 (ISBN)
Public defence
2011-11-25, Hörsal P1, Örebro universitet, Örebro, 09:00 (Swedish)
Opponent
Supervisors
Available from: 2011-10-06 Created: 2011-10-06 Last updated: 2017-10-17Bibliographically approved

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Karlsson, Christina

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