Molecular and clinicopathological analysis of dermatofibrosarcoma protuberans.

Dermatofibrosarcoma protuberans (DFSP) is a dermal and subcutaneous tumor of intermediate malignancy. The most remarkable cytogenetic feature of DFSP is the chromosomal translocation t(17;22)(q22;q13), causing a fusion of the platelet-derived growth factor beta chain (PDGFB) gene at 22q13, and the collagen type 1 alpha 1 (COL1A1) at 17q22. The aim of the study was to analyze the molecular characteristic of DFSP in conjunction with histopathological and clinical features. We performed fluorescence in situ hybridization (FISH) and multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) to detect chromosomal translocations and fusion gene transcripts in 16 formalin-fixed, paraffin-embedded DFSP samples. In addition, the amplification of PDGFB was also evaluated in the 16 DFSP samples by real-time PCR. FISH analysis revealed that all the 16 samples exhibited COL1A1-PDGFB gene fusion. Eleven out of 11 informative cases (100%) showed fusion transcripts by multiplex RT-PCR analysis. Various exons of the COL1A1 gene were fused with the PDGFB gene. Among them, exon 25 was found to be more frequently involved. Real-time PCR showed that the PDGFB copy number increase in the DFSP samples was higher than in normal skin tissues (p=0.007). Values of FISH fusion signals and PDGFB DNA analysis were variable between samples, but suggested that increased values might be associated with parameters of tumor progression. Our results confirm that analysis of the COL1A1-PDGFB status by FISH and RT-PCR is a useful tool in the confirmation of a DFSP diagnosis. In addition, the analysis of PDGFB copy number status may become a useful diagnostic marker since the gene is a potential target for treatment of DFSP patients.

Bone marrow biopsies of patients with hematopoietic and lymphoid disorders - epidemiology, chromosomal aberrations and molecular pathology.

Bone marrow biopsy of the iliac crest is the first and most important step in the diagnostics of hematopoietic disorders. The biopsies of the years 2006 and 2007 from the Institute of Pathology of the Jena University Hospital were retrospectively analyzed for clinicopathological parameters. In addition, the Mitelman database was retrieved for chromosomal aberrations. The analysis of 2820 reports from 1185 patients revealed that lymphomas, plasma cell myeloma and acute leukemia were most frequent. Males predominated in myeloproliferative neoplasms and lymphoma subtypes, particularly CLL, except for plasma cell myeloma and acute leukemia. A peak incidence was seen between 61 and 70 years of age with a varying pattern for single entities. The database search revealed that ALL, AML, CLL and CML were mainly diploid while Hodgkin lymphoma, mature B-cell lymphoma and multiple myeloma mostly carried hyperdiploid chromosome numbers. Numerical aberrations like chromosome 8 gains in hyperdiploid CML were prominent in specific subgroups. Molecular testing is exemplified in CML, plasma cell myeloma and hairy cell leukemia. The study highlights typical clinicopathological characteristics and new genetic findings in hematopoietic and lymphoid neoplasms with relevance for the new WHO classification and beyond. We hope that it may help in the differential diagnosis of bone marrow biopsies.

Identification of biomarkers to distinguish clear cell sarcoma from malignant melanoma.

Clear cell sarcoma is a rare and malignant soft tissue tumor that shows phenotypic and immunohistochemical overlap with cutaneous malignant melanoma; identification of biomarkers that differentiate clear cell sarcoma from malignant melanoma is therefore needed. In this study, we performed mutation analysis of BRAF and NRAS, investigated the EWSR1 gene rearrangement and evaluated the protein expression of insulin-like growth factor 2 and insulin-like growth factor 1R in 31 cases of malignant melanoma and 16 cases of clear cell sarcoma. By direct sequencing and high-resolution melting analysis, we identified BRAF and NRAS mutations in 51.6% and 12.9% of malignant melanoma cases, respectively, while none of clear cell sarcoma harbored BRAF or NRAS mutations. Fluorescence in situ hybridization showed that 78.6% of clear cell sarcoma exhibited the t(12;22)(q13;q12) translocation. The presence of type 1, 2, and 3 EWSR1/ATF1 fusion gene transcripts was confirmed by reverse transcriptase polymerase chain reaction analysis, but type 4 and EWSR1/CREB1 fusion gene transcripts were not found. No fusion transcript could be detected in any of the malignant melanoma cases. Additionally, immunohistochemistry showed that the majority of clear cell sarcoma and malignant melanoma had insulin-like growth factor 2 and insulin-like growth factor receptor 1 expression; however the expression of insulin-like growth factor 1R was significantly higher in clear cell sarcoma compared to melanoma (p = .006). Our results suggest that the combination of BRAF and NRAS mutation analysis with fusion gene detection contributes to diagnosis of malignant melanoma and clear cell sarcoma, and that insulin-like growth factor 1R might be a novel target for the treatment of these two malignancies.

A quantitative co-localization analysis of large unspliced tenascin-C(L) and laminin-5/gamma2-chain in basement membranes of oral squamous cell carcinoma by confocal laser scanning microscopy.

BACKGROUND: A structural interaction of the oncofetal large tenascin-C splice variants (Tn-C(L)) and the gamma2-chain of laminin-5 (Ln-5/gamma2) was recently demonstrated in oral squamous cell carcinoma (OSCC). In situ different patterns of co-localization and co-deposition of both proteins could be detected. Especially the co-localization in re-established basement membrane (BM) structures seemed to be biologically meaningful within the process of tumour progression. METHODS: The amount of Tn-C(L) incorporated in reorganized OSCC BM structures at the tumour margins was investigated by a laser scanning microscopy-based quantitative co-localization analysis. RESULTS: In the BM of normal oral mucosa no Tn-C(L) could be detected. In dysplastic and neoplastic oral mucosa a distinct co-localization of Tn-C(L) and Ln-5/gamma2 in the BM region could be observed. The extent of Tn-C(L) arrangement into reorganized BM structures correlated with malignancy grade. CONCLUSIONS: The results suggest at first, a modulation of carcinomatous BM structures by the inclusion of oncofetal matrix proteins during tumour progression and secondly, the BM incorporation of the adhesion-modulating molecule Tn-C(L) as a pre-invasive structural phenomenon in OSCC.