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

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Methods. HE-gefärbte Schnittpräparate und pathologische Befunde von 42 archivierten Hysterektomie-Präparaten von Tumorfällen mit der Diagnose eines ZAK, wurden bezüglich zervikaler Vorläuferläsionen (ZVL) und Befall des unteren Uterinsegments (UUS) reevaluiert. An ZVL−/UUS+ Fällen wurde eine PCR-Analyse für HPV-DNA durchgeführt. Fälle mit ZVL+/− und UUS-, und UUS+/ZVL+ Fälle, sowie UUS+/ZVL−/HPV+Fälle wurden als primäre ZAK klassifiziert. UUS+/ ZVL−/HPV-Fälle wurden als UUSK reklassifiziert. Repräsentative Tumorschnitte wurden mittels standardisierter, automatisierter Methoden für MLH1, MSH2, PMS2 und MSH6 gefärbt. Vollständig fehlende Expression eines MMRP in Tumorzellen bei positiver interner Kontrolle im Normalgewebe wurde als Verlust der Proteinexpression gewertet. Expressionsprofile primärer ZAK und UUSK wurden verglichen und korreliert (Fisher-Exakt-Test). Zusätzlich wurden alle Tumoren für ER, Vimentin, CEA und p16 gefärbt. Results. 37 Fälle wurden als primäre ZAK klassifiziert (37/42; 88.1%), davon waren 27 UUS−, 8 UUS+/ZVL+ und zwei UUS+/ZVL−/HPV+- Fälle. Fünf UUS+/ZVL−/HPV−- Fälle (5/42; 11.9%) wurden als UUSK reklassifiziert. Die MMRP Expression war in sämtlichen primären ZAK erhalten (37/37; 100%). In vier von fünf UUSK (4/5; 80%) zeigte sich ein Expressionsverlust von mindestens einem MMRP (p
Abstracts Methods. Detailed clinical and histopathologic review of a clinical case and review of the literature using PUBMED. Results. We report on a 70-year-old woman with FIGO Stage 1A Grade II (pT1a pN0 L0 V0 R0) endometrial adenocarcinoma who presented 2 years following total abdominal hysterectomy, bilateral salpingo-oophorectomy and lymphonodectomy with left radius bone metastasis. A left forearm amputation was performed. The tumor measured 7.1×6.9×6.5 cm. The histological diagnosis was a poorly differentiated (G3), endometrioid endometrial carcinoma. Both carcinomas were oestrogen and progesterone receptor negative, vimentin, OSCAR and AE1/AE3 positive. Immunohistochemical stains for S-100, desmin, a-smooth muscle actin and CD 34 were negative. Conclusions. This case highlights the rare and unusual presentation of endometrial cancer. For this reason a review of the literature is also provided for all cases with evidence of bone metastasis at presentation of the disease and as a recurrence. Its diagnosis requires immunohistochemistry and awareness of its possible existence. Poster: Molekularpathologie I SA-P-035 Identification of uncommon PIK3CA-mutations in lung cancer by using pyrosequencing V . Schildgen1 , J . Lüsebrink 1 , J .D . Appel1 , C . Wübben1 , W . Engel-Riedel1 , E . Stoelben1 , O . Schildgen1 , M . Brockmann1 1University Witten/Herdecke, Kliniken der Stadt Köln gGmbH, Köln Aims. Phospatidylinositol-3-kinases (PI3K) play an important role in various cell processes. Oncogenic mutations in the PIK3CA gene which codes for the catalytic subunit have been identified in various malignancies and activate the PI3K/AKT/mTOR pathway which is a critical driver of tumorigenesis. Methods. We tested 41 tumor samples with known KRAS, BRAF, and EGFR mutation status for the occurrence of mutations in the PIK3CA gene using a new commercial pyrosequencing kit. Results. Pyrosequencing revealed 2 mutations (4.9%) in the PIK3CA gene, one in exon 9 and one in exon 20. Both mutations have not yet been identified in lung tumor tissue. Conclusions. The screening of our small patient cohort by pyrosequencing identified two mutations (4.8%) in PIK3CA, on in exon 9 (Q546H) and on in exon 20 (M1043T). Both mutations have not yet been described in lung tumours and seem to be rather uncommon mutations. Future screening of large patient cohorts with pyrosequencing may contribute to the detection of more mutations in lung cancer due to the low limit of detections of this method and may contribute to a better understanding of the interaction of mutations and tumorigenesis. SA-P-036 Detection of EML4-ALK fusion oncogenes in NSCLC using a commercial three-color FISH assay (ZytoLight SPEC TriCheck) B . Schmitt 1 , H . Schultz1 , F . Stellmacher1 , E . Vollmer 1 , T . Goldmann1 1Borstel Research Center, Clinical & Experimental Pathology, Borstel Aims. About 5% of non-small cell lung cancers (NSCLC) harbor an inversion of the short arm of chromosome 2 that causes a rearrangement of the ALK and EML4 genes. The resulting fusion oncoprotein comprises a constitutively active tyrosine kinase. Those patients will benefit substantially from treatment with crizotinib, a Met-/tyrosine kinase inhibitor, highlighting the need for appropriate tests allowing one to reliably detect such fusions in biopsy specimens. Existing techniques such as immunohistochemistry, multiplex PCR, or 2-color FISH using break apart-probes each possess specific strengths and weaknesses. By design, a novel 150 | Der Pathologe · Supplement 1 · 2012 commercial 3-color FISH assay offers several theoretical advantages. However, its actual performance in research and diagnostic settings has not been tested. Methods. We analyzed formalin-fixed and paraffin embedded (FFPE) tissue sections (pulmonary adenocarcinomas) and tissue microarrays (TMA; assembled from adenocarcinomas, squamous and large cell carcinomas) for EML4-ALK rearrangements by 3-color FISH assay using ZytoLight SPEC TriCheck probes (Zytovision). Stained slides were evaluated multiply by pathologists and molecular biologists according to standardized criteria. Cancers exhibiting a characteristic fluorescence pattern in at least 15% of tumor cells were taken as EML4-ALK fusion positive. Results. High quality signals were obtained in FFPE tissues, in individual sections as well as in TMAs. Interobserver variability was negligible for wild type identification. Discrimination between FISH signals from wild type vs. EML4-ALK fusions in cancer tissue was clear and binary owing to probe design, and without the problematic intermediate results observed with break apart-probes in case of a small split. Evaluation was more laborious and less reproducible, however, when more non-tumor tissue was interspersed and tumor cells were harder to identify. Assay hands-on time and total turnover time were less than for multiplex PCR. Conclusions. Detecting EML4-ALK fusions based on a 3-color FISH assay using the “ZytoLight SPEC TriCheck”-probes requires specialized equipment, familiarization of the observer and reliable distinction of tumor cells from non-tumour cells. The approach does not require assessment by multiple observers, unlike assays using break-apart probes. Overall, this approach appears particularly suited for clinical studies and basic research in that it will also identify rare and putative novel fusion variants, as well as deletions and amplifications. SA-P-037 Systematic quantitative cross-validation and content analysis of the 450k methylation array from Illumina J . Rößler1 , O . Ammerpohl2 , J . Gutwein2 , U . Lehmann1 1 2 Medical School Hannover, Institute of Pathology, Hannover, University Hospital Schleswig-Holstein, Institute for Human Genetics, Kiel Aims. The relationship between the beta-values provided by the newly released 450k methylation array from Illumina for individual CpG dinucleotides was compared with quantitative methylation levels obtained by conventional pyrosequencing. In addition, the representation on the Illumina array of two groups of genes, which are important in tumour biology and display extensive aberrations in DNA methylation in cancer (microRNAs and imprinted loci), was assessed in detail. Methods. High molecular weight DNA was isolated from histologically examined 18 fresh-frozen human breast cancer specimens, 4 normal mammary epithelial fractions and 4 human breast cancer cell lines (IPH-926, HCC1937, MDA-MB-134, PCM42) using standard procedures. DNA was bisulfite treated and analyzed on 450k arrays following the manufacturer’s protocol. For primary data processing and analysis the software provided by Illumina was employed. These analyses were complemented and extended by employing the Omics Explorer from Qlucore and the R package IMA. The beta-values for individual loci were cross-validated using conventional pyrosequencing of the same DNA samples independently treated with sodium bisulfite. Results. The newly released 450k array methylation array from Illumina shows a high concordance with quantitative pyrosequencing if identical CpG sites are analysed by the two different methods in cell lines (Spearman r=0.88, p
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Inhalt Der Pathologe · Supplement
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Inhalt Der Pathologe · Supplement
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Editorial Liebe Kolleginnen und Kol
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Kolorektales Karzinom 2 VO-005 Tran
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Keynote Lecture VO-014 Genetic dete
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(AMACR, FASN, GOLM1, GSP-pi, ERG) t
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to assess the correct rate of R1 re
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DNA damage, and cytotoxic drugs. Au
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HCV-positive formalin-fixed and par
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well as MET activation were examine
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or BRAF mutation, c-MYC and SIRT1 e
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AG Pneumopathologie III DO-032 Remo
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immature granulopoiesis showed a st
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ned, if well-defined mantle zones,
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phomas). Most prominent gains or am
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DO-062 Tumor-associated macrophages
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DO-080 DOG1: an immunohistochemical
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AG Oralpathologie DO-087 Detection
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DO-094 Do activated fibroblasts inf
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DO-101 Histopathological analysis o
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DO-108 Identification of potential
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Conclusions. In conclusion, 454 par
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subgroup of patients with B-Raf mut
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DO-002b Impact of terminologies in
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Methods. We performed MCPyV-FISH of
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FR-013 HPV-genotype distribution in
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Methods. Three different techniques
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(i.e. investment in equipment and e
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FR-032 COLD-PCR: a powerful tool in
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dissection (MBLND) technique to imp
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SO-005 Prevalence of mutations in s
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SO-011 Methylation profiling and in
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more effective than SAHA alone and
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SO-022 Specialized pathology review
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SO-027 Ki-67 in mitotic score group
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nificantly associated with advanced
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liver did not correlate with the mu
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AG Paidopathologie II SO-046 Overex
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Results. Histomorphology of the ova
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p42 and p27 have not yet been inves
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SO-068 Recent advances in understan
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SO-075 A novel, dual role of CCN3 i
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Pancreatic Adenocarcinoma SG-137 Se
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sease and 30 colon cancer serum sam
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Conclusions. Although CRC is charac
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differentiated and TNM stage III or
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pressed moderate levels of the prot
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FR-P-037 MALDI imaging reveals COX7
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in the context of therapy response
Page 100 and 101: on primary cells of wild-type and c
Page 102 and 103: chemotherapy was recommended and co
Page 104 and 105: aims are twofold: 1) to verify the
Page 106 and 107: Results. Her2/neu status in TMAs an
Page 108 and 109: prognostic impact was found in rect
Page 110 and 111: provided information on survival ti
Page 112 and 113: Methods. Biopsy specimens from 430
Page 114 and 115: the OS rate was 53% for patients wi
Page 116 and 117: FR-P-098 Immunohistological stainin
Page 118 and 119: FR-P-104 Histopathological evaluati
Page 120 and 121: within the earliest morphologic det
Page 122 and 123: Conclusions. In primary imaging a g
Page 124 and 125: fibrotic neointma development as we
Page 126 and 127: FR-P-129 Alpha-1-antitrypsin-PiZ-an
Page 128 and 129: FR-P-136 The expression of central
Page 130 and 131: cosa and in the periarterial tissue
Page 132 and 133: FR-P-149 Combination of Castleman d
Page 134 and 135: or without different concentrations
Page 136 and 137: Results. In 18 cases (79%) the caus
Page 138 and 139: FR-P-168 Autopsy findings in a 2-ye
Page 140 and 141: FR-P-174 MPGN-like glomerulonephrit
Page 142 and 143: Poster: Gynäkopathologie und Mamma
Page 144 and 145: SA-P-011 Genetic aberrations of pre
Page 146 and 147: Mechanismen, die eine Progression d
Page 148 and 149: internet server. A network of inter
Page 152 and 153: Conclusions. The newly released 450
Page 154 and 155: and desmoplastic reaction are diffe
Page 156 and 157: one patient revealed more than 9 mi
Page 158 and 159: situation, the V600E mutation in th
Page 160 and 161: SA-P-064 Evolution of molecular pat
Page 162 and 163: nic markers such as myf4 and MyoD1
Page 164 and 165: Conclusions. To our knowledge, this
Page 166 and 167: SA-P-085 Expression of the eukaryot
Page 168 and 169: Results. The papillary RCC type II
Page 170 and 171: SA-P-098 Male infertility: assessme
Page 172 and 173: SA-P-104 Process oriented scientifi
Page 174 and 175: Conclusions. The IBDW offers a uniq
Page 176 and 177: L Lasitschka F. DO-046 Lehmann A. F
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