A newly characterized human well-differentiated liposarcoma cell line contains amplifications of the 12q12-21 and 10p11-14 regionsPedeutour, Florence; Maire, Georges; Pierron, Anne; Thomas, David; Garsed, Dale; Bianchini, Laurence; Duranton-Tanneur, Valérie; Cortes-Maurel, Annabelle; Italiano, Antoine; Squire, Jeremy; Coindre, Jean-Michel
doi: 10.1007/s00428-012-1256-5pmid: 22678079
While surgery is the usual treatment for localized well-differentiated and dedifferentiated liposarcomas (WDLPS/DDLPS), the therapeutic options for patients with advanced disease are limited. The classical antimitotic treatments are most often inefficient. The establishment of genetically characterized cell lines is therefore crucial for providing in vitro models for novel targeted therapies. We have used spectral karyotyping, fluorescence in situ hybridization with whole chromosome painting and locus-specific probes, and array-comparative genomic hybridization to identify the chromosomal and molecular alterations of a novel cell line established from a recurring sclerosing WDLPS. The karyotype was hypertriploid and showed multiple structural anomalies. All cells retained the presence of a giant marker chromosome that had been previously identified in the primary cell cultures. This giant chromosome contained high-level amplification of chromosomal regions 12q13-21 and lacked the alpha-satellite centromeric sequences associated with WDLPS/DDLPS. The 12q amplicon was large, containing 370 amplified genes. The DNA copy number ranged from 3 to 57. The highest levels of amplification were observed at 12q14.3 for GNS, WIF1, and HMGA2. We analyzed the mRNA expression status by real-time reverse transcription polymerase chain reaction for six genes from this amplicon: MDM2, HMGA2, CDK4, TSPAN31, WIF1, and YEATS4. mRNA overexpression was correlated with genomic amplification. A second amplicon originating from 10p11-14 was also present in the giant marker chromosome. The 10p amplicon contained 62 genes, including oncogenes such as MLLT10, previously described in chimeric fusion with MLL in leukemias, NEBL, and BMI1.
FGFR1 expression and gene copy numbers in human lung cancerKohler, Lukas; Mireskandari, Masoud; Knösel, Thomas; Altendorf-Hofmann, Annelore; Kunze, Almut; Schmidt, Andreas; Presselt, Norbert; Chen, Yuan; Petersen, Iver
doi: 10.1007/s00428-012-1250-ypmid: 22648708
FGFR1 is a receptor tyrosine kinase of which the ligands belong to the fibroblast growth factor family. To evaluate the significance of FGFR1 in lung cancer, we analysed tumours by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Tissue microarrays were constructed containing 380 lung cancer samples including squamous cell carcinomas (SCC), adenocarcinomas (ADC), non-small cell lung cancer not otherwise specified, metastases, neuroendocrine tumours, large cell lung cancer and small cell lung cancer. FGFR1 expression was analysed by IHC and scored semi-quantitatively by a four-tier approach (0, 1, 2, 3). Using dual-colour interphase FISH with probes specific for the locus on 8p12 and the centromere of chromosome 8 (CEN8), copy numbers of FGFR1 were determined. High expression of FGFR1 was associated with increased FGFR1 gene copy numbers in squamous cell carcinoma (p < 0.001). The FGFR1 locus was equally affected by copy number losses and gains. The higher FGFR1 gene copy numbers in SCC compared to ADC did not reach statistical significance. High copy number amplification of FGFR1 was a very rare event, the FGFR1/CEN8 signal ratio reaching a maximum value of 2.75. There were no significant associations between FGFR1 and clinicopathological parameters. Fibroblast growth factor signalling represents an interesting therapeutic target in lung cancer. However, the pathways are complex with potential oncogenic and anti-oncogenic activities. Our data may help to define criteria for selecting patients that may benefit from these new therapeutic options.
Structural requirements of research tissue banks derived from standardized project surveillanceHerpel, E.; Koleganova, N.; Schreiber, B.; Walter, B.; Kalle, C.; Schirmacher, P.
doi: 10.1007/s00428-012-1258-3pmid: 22653529
Tissue banks constitute decisive and rate-limiting resource and technology platforms for basic and translational biomedical research, notably in the area of cancer. Thus, it is essential to plan and structure tissue banking and allocate resources according to research needs, but essential requirements are still incompletely defined. The tissue bank of the National Center of Tumor Diseases Heidelberg (NCT) was founded with the intention to provide tissues of optimal quality and to prioritize the realization of research projects. We analysed its structure and prospective project management registration as well as tracking records for all projects of the NCT tissue bank as of its start in 2005 in order to obtain information that may be relevant for tissue bank planning. All project proposals submitted to the NCT tissue bank (n = 681) were included in the study. For a detailed evaluation of provided services, only projects that were completed until July 2011 (n = 605) were analysed. For these 605 projects, NCT tissue bank provided 769 specific services. In all projects/services, we recorded project leader, type and amount of material provided, type of research (basic/translational), work load of project and project completion. Furthermore, all completed projects were tracked after 90 days according to a standard protocol to determine principal investigators’ (PI) satisfaction and quality of the provided material. Until July 2011, 605 projects had been successfully completed as documented by material transfer agreement. Of the projects, 72.7 % addressed basic research, 22.3 % were translational research projects and 3 % concerned epidemiological research; 91 % (n = 546) concerned a single PI and the NTC tissue bank. For these projects, 769 specific services were provided. Of these services, 288 concerned providing formalin-fixed and paraffin-embedded (FFPE) tissue (extracts, full size sections), 126 providing fresh frozen materials (including fresh frozen sections), 137 providing tissue micro-array (TMA)-based sections and 199 providing immunohistochemical services. Project tracking demonstrated that all projects had started within 90 days after reception of the material by the PIs, and PI satisfaction with provided material exceeded 97 %. Standardized registration and tracking provides valuable structural information for planning and financing of tissue banks and allocation of resources. The high number of completed projects as well as high user satisfaction demonstrates that structuring of tissue banks should be preferably research-oriented and highly efficient. The comparable number of requests for FFPE and fresh frozen tissue as well as TMA-based services underpins the need for a broad approach in terms of methods and material types in order to fulfil research needs.
Co-expression of VEGF and CA9 in ovarian high-grade serous carcinoma and relationship to survivalWilliams, Emma; Martin, Stewart; Moss, Robert; Durrant, Lindy; Deen, Suha
doi: 10.1007/s00428-012-1252-9pmid: 22699808
Poor prognosis in ovarian high-grade serous carcinoma (HGSC) is largely related to resistance to chemotherapy. Tumour hypoxia is known to be associated with chemotherapy resistance. Stabilisation of hypoxia-inducible factor-1α upregulates the expression of downstream genes such as carbonic anhydrase 9 (CA9) and vascular endothelial growth factor (VEGF). This study was undertaken to analyse the hypoxia profile as indicated by the co-expression of VEGF and CA9 and its correlation with survival. VEGF and CA9 expressions were examined in tissue microarray of 97 cases of ovarian high-grade serous carcinoma using immunohistochemistry. High expression of either VEGF or CA9, individually, was associated with decreased overall survival (p = 0.006 and p = 0.05 respectively). Combined high expression of both markers, to give a ‘hypoxia profile’, was associated with chemotherapy resistance (p = 0.036) and showed worse overall survival with a significant p value (p = 0.001). Using multivariate analysis, hypoxia profile was an independent prognostic factor for overall survival (p = 0.028). The combined high expression CA9 and VEGF phenotype, described as high hypoxia profile group, was significantly associated with increased resistance to chemotherapy and poor overall survival. This group may benefit from combined targeted therapy for effective response in ovarian HGSC.
Histological examination and evaluation of donor bile ducts received during orthotopic liver transplantation—a morphological clue to ischemic-type biliary lesion?Hansen, Torsten; Hollemann, David; Pitton, Michael; Heise, Michael; Hoppe-Lotichius, Maria; Schuchmann, Marcus; Kirkpatrick, C.; Otto, Gerd
doi: 10.1007/s00428-012-1245-8pmid: 22588496
Ischemic-type biliary lesions (ITBL) belong to a group of biliary disorders that are regarded as the major complication in patients with orthotopic liver transplantation (OLT). We performed histological evaluation of donor common bile ducts received during OLT to find morphological clues to the pathomechanisms of ITBL. We investigated 93 grafts of 92 patients (recipients: mean age, 56.5 years; underlying disease: hepatocellular carcinoma (n = 45), alcoholic cirrhosis (n = 16), viral hepatitis with cirrhosis (n = 9), retransplantations (n = 9), others (n = 14); donors: mean age, 53.2 years). Donor common bile ducts were received after recirculation of the hepatic artery prior to biliary end-to-end anastomosis and routinely processed. Statistical evaluation was performed by chi-square analysis and multivariate analysis using a logistic regression model. With regard to ITBL (observed in 19.4 %), the following phenomena were found to be statistically relevant: necrosis of the bile duct wall, arteriolonecrosis, vascular lesions (such as subintimal edema), and intramural bleeding (P < 0.001, P < 0.001, P = 0.029, and P = 0.031, respectively). In logistic regression analysis, arteriolonecrosis was the only parameter with significance (P = 0.001). Based on these results on the morphology of the donor common bile duct, we conclude that these phenomena of vascular damage, reflecting microangiopathy, play a major role in the pathogenesis of ITBL.
In pulmonary sclerosing hemangioma expression of β-catenin, Axin, and C-myc differs between the two cell typesLin, Xu-Yong; Zhang, Di; Zhang, Yong; Fan, Chui-Feng; Dai, Shun-Dong; Wang, En-Hua
doi: 10.1007/s00428-012-1247-6pmid: 22614067
Previous studies have shown that pulmonary sclerosing hemangioma is a tumor derived from primitive respiratory epithelium, but its character and the differentiation status of the two cell types (polygonal and cuboidal) composing the lesion are still controversial. We hypothesize that the polygonal cells are immature compared with cuboidal cells and have higher proliferative activity. To further study this question, we examined the expression of β-catenin, Axin, and C-myc by immunostaining in 45 primary sclerosing hemangioma (PSH) specimens. The two cell types were captured by laser capture microdissection from 28 PSH specimens, and total RNA was extracted. Messenger RNA (mRNA) expression of Axin and C-myc was examined by reverse transcription polymerase chain reaction (RT-PCR). By immunostaining, β-catenin was predominantly strongly expressed on the cell membrane of cuboidal cells, while in polygonal cells, β-catenin was predominantly expressed in the cytoplasm and significantly decreased on cell membranes. Axin was expressed in cuboidal cells in 93 % of our 45 cases, but only expressed in 18 % of these in polygonal cells. C-myc expression in polygonal cells was significantly stronger than in cuboidal cells (P < 0.05). RT-PCR showed that the expression level of Axin mRNA in cuboidal cells was significantly higher than in polygonal cells (P < 0.05), and expression level of C-myc mRNA in polygonal cells was significantly higher than in cuboidal cells (P < 0.05).The two PHS cell types have distinct expression of β-catenin, Axin, and C-myc, suggesting that their differentiation status may be different. The higher expression of C-myc in polygonal cells suggests that these cells might have higher proliferative activity.