title: Tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) is an effective method to analyze genetic aberrations in invasive tumors.
abstract: We amplified various amounts of DNA derived from frozen SF210 and U251NCI human glioblastoma cells, carried out comparative genomic hybridization (CGH) using degenerate oligonucleotide primed-PCR (DOP-PCR) products as test probes, and compared results to analyses performed with probes prepared by standard nick translation. Next we extracted DNA from hematoxylin-eosin (HE)- and methyl green (MG)-stained, microdissected sections of formalin-fixed and paraffin-embedded U251NCI cells, amplified and labeled it by DOP-PCR, and subjected it to CGH. Finally, we used the same methods in multiple samples from a single human mixed glioma tissue. DOP-PCR products from 50 pg to 250 ng of DNA were equally effective in generating the same CGH profiles as the standard method. DOP-PCR products from microdissected pieces of MG-stained cells were effective probes for CGH, but HE-stained samples were not desirable. As the proportion of HE-stained sample increased, CGH profiles deteriorated. DOP-PCR products from microdissected pieces of MG-stained paraffin sections of glioma tissue produced CGH profiles compatible with their histological features. CGH performed with DOP-PCR products from microdissected paraffin blocks allows for the accurate investigation of the cytogenetic characteristics from invasive tumors and of cytogenetic heterogeneity within neoplastic tissue.
authors.affiliation: Department of Neurological Surgery, University of California-San Francisco, San Francisco, California 94143-0808, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Hirose
authors.forename: K
authors.initials: K
authors.lastname: Aldape
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.forename: M S
authors.initials: MS
authors.lastname: Berger
authors.forename: B G
authors.initials: BG
authors.lastname: Feuerstein
mesh_terms: D001254:Astrocytoma / Q000235:genetics / Q000473:pathology; D001932:Brain Neoplasms / Q000235:genetics / Q000473:pathology; D002869:Chromosome Aberrations; D004247:DNA / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D004247:DNA