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

FR-P-136
The expression of central cell cycle regulators in non-small cell
lung cancer (NSCLC) has therapy dependent prognostic impact
J . Cortis 1 , A . Warth 1 , M . Meister 2 , T . Muley 2 , H . Hoffmann 3 , A . Stenzinger 1 ,
P .A . Schnabel 1 , P . Schirmacher 1 , W . Weichert 1
1 University Hospital Heidelberg, Institute of Pathology, Heidelberg,
2 Thoracic Hospital Heidelberg, Heidelberg, 3 Thoracic Hospital Heidelberg,
Department of Thoracic Surgery, Heidelberg
Aims. The relevance of function and expression of central cell cycle regulators
in NSCLC has been discussed for years with controversial and
non-conclusive results. To address this topic, we evaluated expression
patterns of central cell cycle regulators in a very large NSCLC cohort
with the goal to ultimately clarify their clinical-prognostic role.
Methods. Expression of the G1-phase cell cycle regulators CDK4, cyclin
D1 and p16 were analysed by immunhistochemical staining and quantitative
evaluation on tissue microarrays comprising 1045 completely
resected NSCLC. The data were correlated with clinical-pathological
factors, patients’ survival and response to therapy.
Results. Loss of expression of CDK4 (p=0.017), cyclin D1 (p=0.001) and
p16 (p=0.039) were associated with higher UICC stages. In general, high
expression of all cell cycle regulators was associated with better clinical
outcome in a slightly variable way, depending on protein and prognostic
parameter investigated [CDK4, overall survival (OS): p=0.457, disease
specific survival (DSS): p=0.110, disease free survival (DFS): p=0.011;
cyclin D1, OS: p=0.037, DSS: p=0.009, DFS: p=0.111; p16, OS: p=0.204,
DSS: p=0.484, DFS: p=0.004]. These effects were prominent in adenocarcinomas,
adenosquamous carcinomas and in pleomorphic carcinomas,
but less pronounced in squamous cell carcinomas. Interestingly, after
stratification for therapy, the positive association of protein overexpression
with survival was only seen in adenocarcinomas without adjuvant
radio- and chemotherapy whereas the association in tumors with adjuvant
irradiation and chemotherapy was switched in the opposite direction.
Conclusions. The expression of CDK4, cyclin D1 and p16 in NSCLC has
prognostic impact depending on therapy. High expression of all three
proteins was associated with improved clinical outcome in completely
resected NSCLC without adjuvant therapy.
FR-P-137
The FUSE-binding proteins (FBPs) represent essential regulators
responsible for tumor cell proliferation, migration and invasion in
non-small cell lung cancer
M . Malz1 , M . Bovet1 , J . Samarin1 , E . Herpel1 , A . Warth1 , S . Singer1 , T . Muley2 ,
M . Meister2 , H . Hoffmann2 , P . Schnabel1 , P . Schirmacher1 , K . Breuhahn1 1University Hospital Heidelberg, Institute of Pathology, Heidelberg,
2University Hospital Heidelberg, Thoracic Hospital, Heidelberg
Aims. The single-strand nucleic acid binding far upstream element (FU-
SE)-binding proteins (FBP)-1, FBP-2, and FBP-3 represent a multifunctional
protein family, regulating transcriptional and post-transcriptional
processes as well as microRNA biogenesis. Elevated expression and
pro-tumorigenic functions of all FBPs have been described for human
liver cancer. Moreover first data indicated that FBP-1 affects microtubule
dynamics through regulation of MT-destabilizing factors in non-small
cell lung cancer (NSCLC). Therefore we aimed to analyze expression and
functional relevance of FBPs in NSCLC.
Methods. The expression of FBPs was analyzed at the transcript (qPCR)
and protein level (Tissue Microarrays [TMA], Western Blotting) in primary
human NSCLC tissue samples. Using gene-specific siRNAs the
expression of FBPs was inhibited in different NSCLC cell lines (Calu-1,
Calu-6, and A549). Functional consequences of reduced protein expression
on viability (MTT-Assay), proliferation (BrdU-Assay), apoptosis
(FACS-Assay; PARP-cleavage), migration (two-dimensional scratch assay),
and invasion (sprouting assay) were analyzed.
Results. The expression of FBP-1 and FBP-2 was significantly elevated in
human NSCLCs (>60%) in comparison to non-tumorous specimens. In
vitro, transient inhibition of FBP-1 in NSCLC cells (Calu-6) was associated
with decreased tumor cell viability (−76%), proliferation (−83%),
and increased apoptosis (2.8-fold). In contrast, transient inhibition of
FBP-2 predominantly reduced tumor cell migration (−62%) and tumor
cell invasion (−81%), suggesting that both FBP isoforms facilitate distinct
tumor-supporting effects. In addition, FBP-2 inhibition increased FBP-1
expression at the transcript and protein level in A549 cells, demonstrating
that FBP-1 may compensate the loss of FBP-2. Accordingly, the FBP-
1/-2 double-knockdown led to a significant reduction of cell viability
(−69%).
Conclusions. In summary, this study provides evidence that overexpression
of FBP-1 and FBP-2 is frequently detectable in NSCLC tissues and
that both proteins are essential factors for tumor growth and NSCLC
cell dissemination. Furthermore FBP-2 negatively regulates FBP-1 expression,
indicating a functional compensation.
FR-P-138
Interobserver variability in the application of the novel IASLC/
ATS/ERS classification of lung adenocarcinomas
A . Warth1 , A . Stenzinger1 , A .-C . von Brünneck2 , B . Goeppert1 , J . Cortis1 ,
I . Petersen3 , P .A . Schnabel1 , W . Weichert1 1University Hospital Heidelberg, Institute for Pathology, Heidelberg,
2Charité University Hospital Berlin, Institute for Pathology,
3University Hospital Jena, Institute for Pathology
Aims. Recently, an international consensus classification for adenocarcinomas
(ADC) of the lung has been published. The cornerstone of this
new classification is the quantification of different growth patterns. However,
data on the reproducibility performance of this classification in
the routine diagnostic setting are lacking.
Methods. We selected 100 constitutive cases of conventional lung ADC
resection specimens from our archives. All tumor slides were classified
independently by five experienced pulmonary pathologists from three
institutions and by two pathologists in training according to the recommendations
of the IASLC/ATS/ERS.
Results. The most frequent predominant pattern in our cohort was solid
(37%), followed by acinar (35%), lepidic (20%), papillary (5%) and micropapillary
(3%). kappa values for the denomination of a predominant
pattern revealed a substantial agreement for pulmonary pathologists
(0.44—0.72) but only fair agreement for pathologists in training (0.38–
0.47). Interobserver variability was significantly higher in cases with higher
slide numbers (p=0.028) and was considerably reduced in a second
evaluation round after the initiation of a training session. Intraobserver
variability was low (kappa=0.79–0.87). Papillary and micropapillary patterns
were the most complicated patterns to evaluate, while evaluation of
lepidic and solid tumor growth was straightforward. The acinar pattern
ranged in between.
Conclusions. Our data imply that the novel classification of pulmonary
ADC is applicable with adequate interobserver variability if performed
by specifically trained pathologists. However, additional efforts are needed
to harmonize the application of this novel and clinically important
classification scheme of pulmonary ADC.
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