96. Jahrestagung der Deutschen Gesellschaft für Pathologie e. V ...

or BRAF mutation, c-MYC and SIRT1 expression was not found increased.
In addition, within the group of serrated lesions with wild type
KRAS and BRAF, a subgroup was characterized by elevated c-MYC and
SIRT1 expression. In these lesions with mostly high grade intraepithelial
neoplasia and carcinomas, nuclear localization of β-catenin suggested
that activation the wnt signalling pathway may mediate the induction of
c-MYC at the transcriptional level.
Conclusions. In summary, we established a link of oncogenic K-Ras and
B-Raf as well as wnt signalling to activation of the c-MYC oncogene and
SIRT1 in the serrated route to colorectal cancer. The elevated expression
levels observed within higher grades of malignancy point to a crucial
function of c-MYC and SIRT1 in the malignant transformation.
AG Pneumopathologie I
DO-025
HOPE-technique improves diagnostics of Bronchoalveolar
Lavage (BAL)
E . Vollmer 1 , S . Marwitz1 , M . Abdullah1 , C . Vock1 , J .S . Fine2 , S . Visvanathan2 ,
K . Gaede1 , H .-P . Hauber1 , P . Zabel1 , T . Goldmann1 1 2 Research Center Borstel, Borstel, Roche, Inflammation Discovery,
United States
Aims. Besides its application in pulmonary routine diagnostics BAL is
a useful tool for scientific investigations. Because of its limitations in
long term storage we explored in this study the utility of a novel fixative
(HOPE, Hepes glutamic acid buffer mediated Organic solvent Protection
Effect) for retrospective and standardized cell analysis also in regard of
immunological and molecular techniques. This study has been performed
in accordance with the 1964 Declaration of Helsinki and its later
amendments.
Methods. BAL samples, obtained by flexible bronchoscopy from patients
with different diseases, were diluted to a standard cell number of
one million cells and incubated in HOPE-fixative as well as in neutral
buffered 4% formalin with a subsequent paraffin-bloc-embedding. In
addition, for addressing RNA preservation, fresh frozen samples were
included. For enhanced/expanded high-throughput analyses of multiple
BAL samples tissue microarrays of paraffin embedded HOPE-BAL were
also produced.
Results. We have shown that HOPE-BAL cells have an excellent morphology;
besides that this technique allows archiving of BAL cells. By
preservation of proteins and nucleic acids it allows the application of immunocytochemistry
as well as a plethora of molecular techniques like in
situ hybridization, quantitative PCR, transcription microarray analysis,
2-D-Gel-Electrophoresis etc. We showed by targeting some exemplary
molecules the power of screening and validating HOPE-BAL for new
biomarkers.
Conclusions. The HOPE-BAL-technique allows long term storage of BAL
cells and is a unique and novel tool for various molecular based applications
in pulmonary medicine. It combines easy handling in the form of
paraffin blocks with almost no limitations in readout techniques thus
being a step forward into enhanced molecular diagnostics and biobanking.
DO-026
Tissue sparing application of the newly proposed IASLC/ATS/
ERS classification of non-small cell lung cancer shows practical
diagnostic and prognostic impact
W . Sterlacci1 , S . Savic2 , T . Schmid3 , M . Fiegl4 , A . Tzankov 2
1 2 Academic Teaching Hospital Feldkirch, Feldkirch, Austria, University Hospital
Basel, Switzerland, 3Medical University Innsbruck, Center of Operative
Medicine, Austria, 4Medical University Innsbruck, Department of Internal
Medicine, Austria
Aims. The histologic subtype of non-small cell lung cancer (NSCLC) determines
treatment strategies and the need for genetic analyses. Since
most NSCLC are diagnosed on small biopsies or cytology specimens, an
accurate but also tissue sparing approach is necessary. To date, consensus
for a general diagnostic algorithm is lacking.
Methods. To test the diagnostic and clinical relevance of the recently published
multidisciplinary guidelines by the International Association for
the Study of Lung Cancer, American Thoracic Society and European Respiratory
Society, we examined 371 surgically resected NSCLC brought
into tissue microarray format as a surrogate for small biopsies. Adenocarcinomas
(ACA) were graded according to architecture considering
lepidic as well, acinary and papillary as moderately and micropapillary
and solid as poorly differentiated.
Results. The antibody panel TTF-1, p63, CK5/6 and CK7 proved diagnostic
for most cases. The positive predictive value (PPV) of p63 and CK5/6
for squamous cell carcinoma (SCC), and of CK7 and TTF1 for ACA was
88.9%, 84.9%, 88.4% and 97.7%, respectively. The negative predictive value
(NPV) of p63 and CK5/6 for SCC, and of CK7 and TTF1 for ACA was
99.5%, 99.5%, 93.6% and 76.9%, respectively. Faint/focal staining for CK7
is negligible for classificatory purposes and focal expression of TTF-1
with variable staining intensity is a feature compatible with SCC (approximately
3% of cases). Overall survival in months for ACA according to
architecture-based tumor grade was 72.5 for well, 71.0 for moderate and
35.7 for poor differentiation (p=0.039).
Conclusions. We propose double stains combining an above mentioned
nuclear and membranous marker, which is highly diagnostic for NSCLC
on small biopsies while conserving tumor tissue for subsequent analyses.
No recommendations using less than 2 sections exist, however a panel
consisting of TTF-1 in combination with CK5/6 may be feasible, since
TTF-1 appears to be the most specific discriminating marker between
ACA and SCC (best PPV for ACA) and the only unequivocally evaluable
staining combination is with cytoplasmic staining for CK5/6, which
also achieved the best NPV for ACA. When grading ACA, the histologic
tumor architecture should be the determining factor. This approach primarily
has prognostic implications but will also result in easier comparisons
of future studies.
DO-027
Multi-immunassay with concurrent staining of 6 antibodies
allows tissue-sparing diagnosis on small tissue samples on nonsmall
cell lung carcinomas with high diagnostic accuracy
G . Kayser1 , A . Csanadi 1 , C . Otto1 , S . Dango2 , B . Passlick2 , M . Werner1 1 2 Institute of Pathology, University Hospital Freiburg, Freiburg, Department
of Thoracic Surgery, University Hospital Freiburg, Freiburg
Aims. Today the histological differentiation of non-small cell lung carcinomas
NSCLC into adenocarcinomas (LAC), squamous cell carcinomas
(SCC), and large cell neuroendocrine carcinomas (LCNEC) is not only of
prognostic relevance but far more of predictive value for different therapeutic
regimes. As these decisions are of utmost relevance in advanced
cancer stages, pathologists are asked to perform a highly accurate diagnosis
on small tissue samples. We here investigated the possibility of simultaneous
staining of widely agreed upon markers for the histological
classification of NSCLC.
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