論文

査読有り
2015年1月

The PFA-AMeX method achieves a good balance between the morphology of tissues and the quality of RNA content in DNA microarray analysis with laser-capture microdissection samples

JOURNAL OF TOXICOLOGIC PATHOLOGY
  • Takeshi Watanabe
  • ,
  • Atsuhiko Kato
  • ,
  • Hiromichi Terashima
  • ,
  • Koichi Matsubara
  • ,
  • Yu Jau Chen
  • ,
  • Kenji Adachi
  • ,
  • Hideaki Mizuno
  • ,
  • Masami Suzuki

28
1
開始ページ
41
終了ページ
47
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1293/tox.28.2014-0045
出版者・発行元
JAPANESE SOC TOXICOLOGIC PATHOLOGY

Recently, large-scale gene expression profiling is often performed using RNA extracted front unfixed frozen or formalin-fixed paraffin embedded (FFPE) samples. However, both types of samples have drawbacks in terms of the morphological preservation and RNA quality. In the present study, we investigated 30 human prostate tissues using the PFA-AMeX method (fixation using paraformaldehyde (PFA) followed by embedding in paraffin by AMeX) with a DNA microarray combined with laser-capture microdissection. Morphologically, in contrast to the case of atypical adenomatous hyperplasia, loss of basal cells in prostate adenocarcinomas was as obvious in PFA-AMeX samples as in FFPE samples. As for quality, the loss of rRNA peaks 18S and 28S on the capillary electropherograms from both FETE and PFA-AMeX samples showed that the RNA was degraded equally during processing. However, qRT-PCR with 3' and 5' primer sets designed against human beta-actin revealed that, although RNA degradation occurred in both methods, it occurred more mildly in the PFA-AMeX samples. In conclusion, the PFA-AMeX method is good with respect to morphology and RNA quality, which makes its promising tool for DNA microarrays combined with laser-capture microdissection, and if the appropriate RNA quality criteria are used, the capture of credible GeneChip data is well over 80% efficient, at least in human prostate specimens.

リンク情報
DOI
https://doi.org/10.1293/tox.28.2014-0045
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/26023261
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000358704000007&DestApp=WOS_CPL
ID情報
  • DOI : 10.1293/tox.28.2014-0045
  • ISSN : 0914-9198
  • eISSN : 1881-915X
  • PubMed ID : 26023261
  • Web of Science ID : WOS:000358704000007

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