50
Chromosome, Microdissection and Microcloning
laser desorption/ionization (SELDI) that
allows the retention of proteins on a
solid-phase chromatographic surface (Pro-
teinChip
Array)
with
direct
detection
of retained proteins by time of flight-
mass spectrometry (TOF-MS). The use
of rapid, high-throughput mass spectro-
metric–based ±ngerprints of peptides and
proteins may prove valuable for new
molecular classi±cation of human tumors
and disease stages. Coupled with laser-
capture microdissection, high-density pro-
tein arrays, antibody arrays, and small
molecular arrays could have a substantial
impact on proteomic pro±ling of human
malignancies.
Differentially expressed proteins in nor-
mal and neoplastic tissues were recently
analyzed by von Eggeling et al. by com-
paring the protein expression patterns
generated using SELDI-based TOF-MS
in cases of renal cell carcinoma, periph-
eral and central tumor tissue, as well
as four microdissected cases of cervi-
cal intraepithelial neoplasia and three
microdissected cases of cervix uteri car-
cinoma. By applying ProteinChip array
technology, it was possible to investi-
gate simultaneous and complex changes
associated with tumor development and
progression at the protein level in sev-
eral cancers. In their landmark article,
Petricoin et al. declared that the ultimate
goal of proteomics is to translate the
bench-side promise into bedside reality.
It is to characterize the information flow
through protein networks. This informa-
tion can be a cause, or a consequence,
of disease processes. Using cancer as
a model disease, clinical proteomics ap-
plications are being developed to detect
cancer earlier, to discover the next genera-
tion of targets and imaging biomarkers,
and ±nally to tailor the therapy to the
patient.
6.8
Incorporation of Microdissection
Techniques in Modern Pathology
The last decade has witnessed a ‘‘molecu-
lar
’revo
lu
t
ioninpa
tho
logy
.Demons
tra
t
-
ing that transcription of speci±c single
genes or small gene sets and their protein
products by
in situ
hybridization and im-
munocytochemistry has become routine
in diagnostic and experimental pathology
laboratories. A perhaps greater revolution
is imminent with the application of more
recently established and emergent tech-
nologies. These include new approaches
to polymerase chain reaction (PCR); si-
multaneous studies of multiple genes and
their expression using oligonucleotide and
cDNA arrays; serial analysis of gene ex-
pression (SAGE); expressed sequence tag
(EST) sequencing, subtractive cloning, and
differential display; high-throughput se-
quencing; comparative genomic hybridiza-
tion, multiplex fluorescence
in situ
hy-
bridization (FISH) (spectral karyotyping);
reverse chromosome painting; and knock-
out and transgenic organisms. Laser mi-
crodissection and micromachining, and
new methods in bioinformatics, ‘‘data
mining,’’ and data visualization are added
to the list. The use of any combina-
tion of these molecular methods will
profoundly change diagnosis, prognosis,
and treatment targeting oncology and
will elucidate fundamental mechanisms
of neoplastic transformation. Individual
susceptibility to speci±c diseases will be-
come assessable and screening will be
re±ned.
Microdissection of chromosomal tran-
slocations,
breakpoints,
HSR,
marker
and ring chromosomes, micronuclei, and
reverse-painting normal metaphase chro-
mosomes have shed light on the origin
of these aberrations. Table 2 presents a
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