Tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) is an effective method to analyze genetic aberrations in invasive tumors

Citation
Y. Hirose et al., Tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) is an effective method to analyze genetic aberrations in invasive tumors, J MOL DIAGN, 3(2), 2001, pp. 62-67
Citations number
26
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Research/Laboratory Medicine & Medical Tecnology
Journal title
JOURNAL OF MOLECULAR DIAGNOSTICS
ISSN journal
1525-1578 → ACNP
Volume
3
Issue
2
Year of publication
2001
Pages
62 - 67
Database
ISI
SICI code
1525-1578(200105)3:2<62:TMADOP>2.0.ZU;2-Q
Abstract
We amplified various amounts of DNA derived from frozen SF210 and U251NCI h uman glioblastoma cells, carried out comparative genomic hybridization (CGH ) using degenerate oligonucleotide primed-PCR (DOP-PCR) products as test pr obes, and compared results to analyses performed with probes prepared by st andard nick translation, Next we extracted DNA from hematoxylin-eosin (HE)- and methyl green (MG)-stained, microdissected sections of formalin-fixed a nd paraffin-embedded U251NCI cells, amplified and labeled it by DOP-PCR, an d subjected it to CGH, Finally, we used the same methods in multiple sample s from a single human mixed glioma tissue. DOP-PCR products from 50 pg to 2 50 ng of DNA were equally effective in generating the same CGH profiles as the standard method. DOP-PCR products from microdissected pieces of MG-stai ned cells were effective probes for CGH, but HE-stained samples were not de sirable, As the proportion of HE-stained sample Increased, CGH profiles det eriorated. DOP-PCR products from microdissected pieces of MG-stained paraff in sections of glioma tissue produced CGH profiles compatible with their hi stological features, CGH performed with DOP-PCR products from microdissecte d paraffin blocks allows for the accurate investigation of the cytogenetic characteristics from invasive tumors and of cytogenetic heterogeneity withi n neoplastic tissue.