Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2006-11-30
pubmed:abstractText
Coupling array technology to laser capture microdissection (LCM) has the potential to yield gene expression profiles of specific cell populations within tissue. However, remaining problems with linear amplification preclude accurate expression profiling when using the low nanogram amounts of RNA recovered after LCM of human tissue. We describe a novel robust method to reliably amplify RNA after LCM, allowing direct probing of 12K gene arrays. The fidelity of amplification was demonstrated by comparing the ability of amplified RNA (aRNA) versus that of native RNA to identify differentially expressed genes between two different cell lines, demonstrating a 99.3% concordance between observations. Array findings were validated by quantitative polymerase chain reaction analysis of a randomly selected subset of 32 genes. Using LCM to recover normal (N=5 subjects) or cancer (N=3) cell populations from intact human prostate tissue, three differentially expressed genes were identified. Independent investigators have previously identified differential expression of two of these three genes, hepsin and beta-microseminoprotein, in prostate cancer. Taken together, the current study demonstrates that accurate gene expression profiling can readily be performed on specific cell populations present within complex tissue. It also demonstrates that this approach efficiently identifies biologically relevant genes.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1365-7852
pubmed:author
pubmed:issnType
Print
pubmed:volume
9
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
379-91
pubmed:dateRevised
2007-12-3
pubmed:meshHeading
pubmed-meshheading:16786039-Cell Line, Tumor, pubmed-meshheading:16786039-Epithelial Cells, pubmed-meshheading:16786039-Gene Expression Profiling, pubmed-meshheading:16786039-Gene Expression Regulation, Neoplastic, pubmed-meshheading:16786039-Humans, pubmed-meshheading:16786039-Male, pubmed-meshheading:16786039-Microdissection, pubmed-meshheading:16786039-Nucleic Acid Amplification Techniques, pubmed-meshheading:16786039-Oligonucleotide Array Sequence Analysis, pubmed-meshheading:16786039-Polymerase Chain Reaction, pubmed-meshheading:16786039-Prostate, pubmed-meshheading:16786039-Prostatic Neoplasms, pubmed-meshheading:16786039-RNA, Neoplasm, pubmed-meshheading:16786039-Reproducibility of Results, pubmed-meshheading:16786039-Transcription, Genetic, pubmed-meshheading:16786039-Tumor Cells, Cultured
pubmed:year
2006
pubmed:articleTitle
Characterization of a method for profiling gene expression in cells recovered from intact human prostate tissue using RNA linear amplification.
pubmed:affiliation
Department of Medicine, Division of Hematology/Oncology, Northwestern University Medical School and the Robert H Lurie Cancer Center of Northwestern University, Chicago, IL, USA.
pubmed:publicationType
Journal Article, Research Support, N.I.H., Extramural, Validation Studies