Publication: Gene expression analysis
in sections and tissue microarrays of
archival tissues by mRNA in situ hybridization
Authors
Henke, R.T. ; Maitra, A. ; Palk, S. ; Wellstein, A.
item.page.secondaryauthor
item.page.director
Publisher
Murcia : F. Hernández
publication.page.editor
publication.page.department
DOI
item.page.type
info:eu-repo/semantics/article
Description
Abstract
Altered expression of genes in diseased
tissues can prognosticate a distinct natural progression of
the disease as well as predict sensitivity or resistance to
particular therapies. Archival tissues from patients with a
known medical history and treatments are an invaluable
resource to validate the utility of candidate genes for
prognosis and prediction of therapy outcomes. However,
stored tissues with associated long-term follow-up
information typically are formalin-fixed, paraffinembedded
specimen and this can severely restrict the
methods applicable for gene expression analysis. We
report here on the utility of tissue microarrays (TMAs)
that use valuable tissues sparingly and provide a
platform for simultaneous analysis of gene expression in
several hundred samples. In particular, we describe a
stable method applicable to mRNA expression screening
in such archival tissues. TMAs are constructed from
sections of small drill cores, taken from tissue blocks of
archival tissues and multiple samples can thus be
arranged on a single microscope slide. We used mRNA
in situ hybridization (ISH) on >500 full sections and
>100 TMAs for >10 different cDNAs that yielded
>10,000 data points. We provide detailed experimental
protocols that can be implemented without major hurdles
in a molecular pathology laboratory and discuss
quantitative analysis and the advantages and limitations
of ISH. We conclude that gene expression analysis in
archival tissues by ISH is reliable and particularly useful
when no protein detection methods are available for a
candidate gene.
publication.page.subject
Citation
item.page.embargo
Ir a Estadísticas
Sin licencia Creative Commons.