TY - JOUR
T1 - Expression profiling of ductal carcinoma in Situ by laser capture microdissection and high-density oligonucleotide arrays
AU - Luzzi, Veronica
AU - Holtschlag, Victoria
AU - Watson, Mark A.
N1 - Funding Information:
Supported in part by grant no. 016-99 from the Mary Kay Ash Charitable Foundation .
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0034965544&partnerID=8YFLogxK
U2 - 10.1016/S0002-9440(10)64672-X
DO - 10.1016/S0002-9440(10)64672-X
M3 - Article
C2 - 11395378
AN - SCOPUS:0034965544
SN - 0002-9440
VL - 158
SP - 2005
EP - 2010
JO - American Journal of Pathology
JF - American Journal of Pathology
IS - 6
ER -