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