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

Methods. Biopsy specimens from 430 untreated patients were investigated
immunohistochemically with monoclonal antibodies against topo
IIα (Ki-S4).
Results. Patients with low (50%) topo IIα expression. The Kaplan-Meier analysis showed
a significant difference in the overall survival time related to the percentage
of topo IIα (p=0.0012) positive tumor cells.
Conclusions. Patients with colorectal cancer and high expression level of
topo IIα have a superior clinical outcome compared to patients with low
expression levels.
FR-P-085
Stem cell marker cancer testis antigen (CT 45) expression in colorectal
cancer: an immunhistochemical study of 704 patients
C . Schrader1 , F . Gieseler2 , J .H . Bräsen3 , B . Sipos4 , C . Röcken3 , W . Klapper3 ,
H . Kalthoff5 , W . v . Schönfels5 , R . Lucius6 , C . Pflüger1 , S . Hinz5 , H . Held7 , T . Raff1 ,
G . Klöppel8 , J . Claasen1 , S . Nazzal1 , C . Dreyer1 , C . Schafmayer5 1 2 2 University Hospital of Kiel, nd Department of Medicine, Kiel, UKSH, Campus
Lübeck, I . Department of Medicine, 3UKSH, Campus Kiel, Department
of Pathology, 4University of Tübingen, Department of Pathology, 5UKSH, Campus Kiel, Department of Surgery, 6UKSH, Campus Kiel, Department of
Anatomy, 7Hospital Neumünster, Department of Medicine, Kiel, 8University Munich, Department of Pathology
Aims. A subset of colorectal cancer derived from stem cells. Cancer testis
antigen (CT45) is expressed in immature gonocytes, spermatogonia and
also in germ cell tumors. There are only limited data about CT 45 expression
in colorectal cancer and no data if the expression in tumor cells has
an influence on the clinical course of the disease.
Methods. We investigated immunohistochemically embedded tissue
from 704 patients with a monoclonal antibody against CT 45 (Ki-A10).
The percentage of CT45 expressing cells was quantified according to the
following classification: negative, 0–25%, 25–50%, 50–75% and more than
75% positive tumor cells.
Results. The majority of cases were negative (n=633). 71 tumor specimens
were CT45 positive. The Kaplan Meier analysis revealed no significant
difference (p=0.11) in the clinical outcome of CT 45 positive and negative
cases
Conclusions. Cancer testis antigen (CT 45) is expressed in a subset of
colorectal cancer. The CT 45 expression is no prognostic factor for the
clinical outcome of the disease, but it might have clinical implication for
therapy decision of stem cell derived cancer.
FR-P-086
SOX9 promotes cell migration and anchorage-independent
growth of colorectal cancer cell lines
C . Hui1 , X . Enping1 1Zhejiang University, School of Medicine, Hangzhou, China
Aims. SOX9 is a high-mobility group (HMG) domain transcription factor
that plays a critical role in multiple aspects of development and differentiation.
There is a close connection between normal development
and oncogenesis. Recent studies have revealed that SOX9 is implicated
in several types of cancer progression, but SOX9 does not have consistent
function roles in different cancers. Importantly, the function and
role of SOX9 in colorectal cancer (CRC) exactly remains unclear. In our
previous study we identified that SOX9 overexpression in CRC as an independent
predictor of an unfavorable outcome for CRC in Chinese population.
This study is to show the role of SOX9 in CRC cell lines.
Methods. Western blotting, RT-PCR and Q-PCR were performed to
detect the expression of SOX9 in CRC cell lines. Stably SOX9-overexpression
RKO cell was constructed. Subsequently, Cell Counting Kit-8
assay and Soft agar colony formation assay, Annexin v-PE/7-AAD Double
Stain Apoptosis Detection assay, xCELLigence system, cell migration
and 5-Fu cell toxicity test were performed to detect the roles of SOX9 in
vitro.
Results. After performing gain-of-function studies, it was clarified that
the SOX9-overexpression RKO cell had no significant effects on cell
proliferation, cell apoptosis and 5-Fu cell toxicity; In migration assay, it
is revealed that SOX9 could promote cell migration by transwell chamber
and xCELLigence system. Also it was assessed that the SOX9-overexpression
RKO cell had impact on anchorage-independent growth by
using soft agar colony formation assay.
Conclusions. The expression of SOX9 can promote cell migration and
anchorage-independent growth, but it has no effect on cell proliferation.
FR-P-087
RegIV promotes migration and invasion of CRC cell lines
Z . Jie1 , W . Jingyu1 , L . Maode1 1Zhejiang University, School of Medicine, Hangzhou, China
Aims. The regenerating gene IV (RegIV) plays an important role in colorectal
tumorigenesis. However, its biological functions have not been
well elucidated.
Methods. RT-PCR and western blotting analysis were performed to detect
the expression of RegIV in colorectal carcinoma (CRC) cell lines
HT29, RKO and LOVO, then the stably RegIV overexpressed and Reg4
shRNA-silenced cell lines were constructed in the RegIV negative-expressed
and positive-expressed cell lines respectively. Subsequently, Cell
counting Kit-8 (cck-8) assay, EdU assay and xCELLigence system assay
were performed to detect cell proliferation. Transwell chamber and matrigel
assays were used to detect cell migration and invasion.
Results. RT-PCR and western blotting showed that RKO and HT29 cell
lines do not expressed RegIV, while LOVO cell lines expressed RegIV.
After performing gain-of-function and loss-of-function studies, it has
been shown that both RegIV overexpressed and RegIV shRNA-silenced
cell lines have no significant effects on cell proliferation. In contrast to
control, migration and invasion abilities are significantly increased on
RegIV overexpressed cell line and are significantly decreased on RegIV
shRNA- silenced cell lines.
Conclusions. The expression of RegIV can promote CRC cell lines to migrate
and invade, but it has no effect on proliferation.
FR-P-088
Minichromosome maintenance protein 6 (MCM 6) expression in
colorectal cancer: a proliferation marker and a prognostic factor
for clinical outcome
C . Schrader1 , F . Gieseler2 , J .H . Bräsen3 , B . Sipos4 , C . Röcken3 , W . Klapper3 ,
H . Kalthoff5 , W . v . Schönfels5 , R . Lucius6 , C . Pflüger1 , S . Hinz5 , H . Held7 , T . Raff1 ,
G . Klöppel8 , J . Claasen1 , S . Nazzal1 , C . Schafmayer5 1 2 2 University Hospital of Kiel, nd Department of Medicine, Kiel, UKSH, Campus
Lübeck, I . Department of Medicine, 3UKSH, Campus Kiel, Department
of Pathology, 4University of Tübingen, Department of Pathology, 5UKSH, Campus Kiel, Department of Surgery, 6UKSH, Campus Kiel, Department of
Anatomy, 7Hospital Neumünster, Department of Medicine, Kiel, 8University Munich, Department of Pathology
Aims. MCM6 is one of six proteins of the MCM family which are involved
in the initiation of DNA replication and is a marker of proliferating
cells. Proliferation markers and their expression levels as prognostic factors
are discussed controversial in colorectal cancer.
Methods. We investigated paraffin embedded tissue from 570 patients
with colorectal cancer with stage 1 to 4 immunohistochemically with
a monoclonal antibody against MCM6. 500 tumor cells were counted
and the percentage of positive cells was calculated. Overall survival time
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