Document Detail


A lung tissue bank for gene expression studies in chronic obstructive pulmonary disease.
MedLine Citation:
PMID:  17136987     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A bank of surgically resected human lung tissues frozen at -70 degrees C after being inflated with support medium for cutting frozen tissue and a separate group inflated with fixative and embedded in paraffin has been established for studies of chronic obstructive pulmonary disease. The present report concerns the quality of RNA that can be extracted from these frozen and fixed tissue samples and from cells obtained from them by laser capture microdissection. The results show that the RNA yield was 257+/-183 ng/mg and 77+/-56 ng/mg from randomly selected frozen and paraffin-embedded tissue, respectively. Intact 18S and 28S rRNA subunits were present in 11/23 frozen and 2/6 paraffin-embedded specimens. The 375-bp actin and 296-bp glyceraldehdye 3-phosphate dehydrogenase targets were amplified by reverse transcription-PCR from both sources and the 983-bp glyceraldehdye 3-phosphate dehydrogenase and 499-bp nonhousekeeping integrin-linked kinase targets from frozen tissue. The minimal amount of RNA required for reverse transcription-PCR of 296-bp glyceraldehdye 3-phosphate dehydrogenase target was 29 pg from frozen tissue when RNA subunits were present and 144 pg when these subunits were absent compared to 0.8 ng from paraffin-embedded tissue. Ten laser pulses were required to laser capture sufficient cells from frozen tissue to detect amplification of the 375-bp actin target while more pulses were required for equivalent amplification from paraffin-embedded tissue. Storage time had no detectable effect on RNA quality. We conclude that both frozen and paraffin-embedded tissues as well as laser-captured cells are suitable for gene expression studies but frozen tissue offered greater sensitivity.
Authors:
Lily Ding; Kevin B Quinlan; W Mark Elliott; Mowafak Hamodat; Peter D Paré; James C Hogg; Shizu Hayashi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  COPD     Volume:  1     ISSN:  1541-2555     ISO Abbreviation:  COPD     Publication Date:  2004  
Date Detail:
Created Date:  2006-12-01     Completed Date:  2006-12-19     Revised Date:  2009-11-17    
Medline Journal Info:
Nlm Unique ID:  101211769     Medline TA:  COPD     Country:  United States    
Other Details:
Languages:  eng     Pagination:  191-204     Citation Subset:  IM    
Affiliation:
Department of Pathology, University of British Columbia iCAPTURE Centre/ McDonald Research Laboratories, Vancouver, British Columbia, Canada.
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MeSH Terms
Descriptor/Qualifier:
Actins / genetics*
Blotting, Southern / methods
Cell Line
Gene Expression*
Glyceraldehyde-3-Phosphate Dehydrogenases / genetics*
Humans
Pulmonary Disease, Chronic Obstructive / genetics*,  pathology
RNA, Messenger / genetics*
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction / methods
Tissue Banks*
Chemical
Reg. No./Substance:
0/Actins; 0/RNA, Messenger; EC 1.2.1.-/Glyceraldehyde-3-Phosphate Dehydrogenases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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