Expression profiling of ductal carcinoma in Situ by laser capture microdissection and high-density oligonucleotide arrays

Veronica Luzzi, Victoria Holtschlag, Mark A. Watson

Research output: Contribution to journalArticlepeer-review

107 Scopus citations

Abstract

Gene expression profiling through the use of nucleic acid arrays is a powerful method for the molecular classification of human neoplasms. Laser capture microdissection is an equally useful technique to selectively isolate defined cell populations from heterogeneous histological tissue sections. In this report, we demonstrate how a modest use of laser capture microdissection is sufficient to isolate nanogram quantities of high-quality RNA. Together with the use of several internal standards and microcapillary electrophoresis of input RNA, two rounds of linear molecular amplification have been used to generate sufficient quantities of labeled target for hybridization to high-density oligonucleotide expression arrays. Results demonstrate that the technique is reproducible, generates only modest biasing of the original transcript population, and is comparable to the sensitivity achieved with standard methodology. Using this approach, we have compared the expression profiles of nonmalignant human breast epithelium and adjacent ductal carcinoma in situ lesions from breast cancer patients. Several genes, previously implicated in human breast cancer progression, demonstrate differential expression among the microdissected cell populations.

Original languageEnglish
Pages (from-to)2005-2010
Number of pages6
JournalAmerican Journal of Pathology
Volume158
Issue number6
DOIs
StatePublished - 2001

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