Expert consensus document:Cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA)

Cholangiocarcinoma (CCA) is a heterogeneous group of malignancies with features of biliary tract differentiation. CCA is the second most common primary liver tumour and the incidence is increasing worldwide. CCA has high mortality owing to its aggressiveness, late diagnosis and refractory nature. In May 2015, the "European Network for the Study of Cholangiocarcinoma" (ENS-CCA: www.enscca.org or www.cholangiocarcinoma.eu) was created to promote and boost international research collaboration on the study of CCA at basic, translational and clinical level. In this Consensus Statement, we aim to provide valuable information on classifications, pathological features, risk factors, cells of origin, genetic and epigenetic modifications and current therapies available for this cancer. Moreover, future directions on basic and clinical investigations and plans for the ENS-CCA are highlighted

Induction of multidrug resistance proteins in lymphocytes from patients with arthritic disorders

[No abstract available

Increased metastatic lymph node 64 and CYP17 expression are associated with high stage prostate cancer

The metastatic lymph node 64 (MLN64), which is localized in the human chromosome 17, encodes a protein with strong homology with steroidogenic acute regulatory protein. Its overexpression in human breast carcinomas and MLNs led to the hypothesis that this protein could be involved in intraneoplastic steroidogenesis. In the present study, we investigated the expression of MLN64 in prostate cancer, another hormone-dependent tumor, and compared its expression with that of CYP17, the gene encoding for the key enzyme of androgen synthesis. We investigated by RT-PCK the expression of MLN64 and CYP17 in 60 prostatic tumors and compared their expression with the stage of disease and the appearance of relapses in a follow-up of 24 months. We found MLN64 and CYP17 expressed in all samples examined, with significantly higher expression neoplastic tissues with respect to normal tissues (NTs). Moreover, only in neoplastic but not in NTs, a positive linear correlation was found between MLN64 and CYP17 gene expression. MLN64 and CYP17 expression seems to correlate with high stage, high Gleason score and short relapse-free time. These data, for the first time, demonstrate the presence of MLN64 and CYP17 expression in both normal and neoplastic prostatic tissues. The biological role of MLN64 in human prostate and, particularly, in neoplastic tissue is still unclear. Our findings concerning MLN64 and CYP17 gene expression and their significant positive correlation in human prostate cancer may suggest their possible role in intraneoplastic autonomous steroidogenesis

Isolation and Characterization of Circulating Tumor Cells in Squamous Cell Carcinoma of the Lung Using a Non-EpCAM-Based Capture Method

<div><p>Introduction</p><p>The exclusion of circulating tumor cells (CTCs) that have lost epithelial antigens during the epithelial-to-mesenchymal transition (EMT) process by using Epithelial Cell Adhesion Molecule (EpCAM) based capture methods is still a matter of debate. In this study, cells obtained after depletion procedure from blood samples of squamous cell lung cancer (SQCLC) patients were identified based on morphology and characterized with the combination of FISH assessment and immunophenotypic profile.</p><p>Materials and Methods</p><p>Five mL blood samples, collected from 55 advanced SQCLC patients, were analyzed by a non-EpCAM-based capture method. After depletion of leukocytes and erythroid cells, the negative fraction was characterized by both FISH using a fibroblast growth factor receptor 1 (FGFR1) probe and by immunocytochemistry. Thirty healthy donors were also tested.</p><p>Results</p><p>Based on morphology (nuclear dimension ≥10 μm, shape and hypercromatic aspect) suspicious circulating cells clearly distinguishable from contaminant leukocytes were observed in 49/55 (89%) SQCLC patients. Thirty-four of the 44 (77%) samples evaluable for FGFR1 FISH showed ≥ 6 FGFR1 gene copy number on average per cell. Vimentin expression involved 43% (18/42) of pooled circulating SQCLC cells, whereas only 29% (14/48) were EpCAM positive. Confocal microscopy confirmed the localization of FGFR1 probe in suspicious circulating cells. Suspicious circulating elements were also observed in healthy donors and did not show any epithelial associated antigens. A significantly lower number of suspicious circulating cells in healthy donors compared to SQCLC patients was found.</p><p>Conclusions</p><p>Among the heterogeneous cell population isolated by depletion procedure, the coexistence of cells with epithelial and/or mesenchymal phenotype suggests that EMT may participate to transendothelial invasion and migration of tumor cells in advanced SQCLC. The finding of cells with neither EpCAM or EMT phenotype, retrieved after non-EpCAM-based systems, underlines the presence of suspicious elements in the blood of both SQCLC patients and healthy donors. Further phenotyping and molecular analyses are necessary to fully characterize these circulating elements.</p></div

Distribution of suspicious circulating elements isolated by depletion procedure from SQCLC patients and healthy donors.

<p>Distribution of suspicious circulating elements isolated by depletion procedure from SQCLC patients and healthy donors.</p

Confocal images of propidium iodide (PI) stained suspicious circulating elements obtained from SQCLC patients.

<p>The upper part of the figure shows a PI stained SCE with multiple green signals referring to multiple copies of <i>FGFR1</i> gene (scale bar = 5 μm); the middle part shows a PI stained nucleus with EpCAM expression (scale bar = 5 μm) and the bottom part a PI stained nucleus with vimentin expression (scale bar = 10 μm) associated with two small leukocytes showing a faint cytoplasmic fluorescence.</p

Antibodies and their dilutions used for immunocytochemistry.

<p>FITC, Fluorescein isothiocyanate.</p><p>Antibodies and their dilutions used for immunocytochemistry.</p

Comparison between depletion procedure and EpCAM-based enrichment system in 25 advanced SQCLC patients.

<p>EpCAM, epithelial cell adhesion molecule.</p><p>Comparison between depletion procedure and EpCAM-based enrichment system in 25 advanced SQCLC patients.</p

Suspicious circulating elements in SQCLC patients.

<p>Representative image of a suspicious circulating element (SCE) isolated by depletion procedure and stained by hematoxylin; contaminant platelets and red blood cell ghosts are present in the background (A). DAPI stained SCE nucleus and small contaminant white blood cells (B). Confocal image of a large hyperchromatic SCE nucleus labelled by the red fluorescence of propidium iodide (PI); a small PI stained nucleus of a white blood cell is also present (C). Weak hematoxylin and eosin stained nucleus of a SCE isolated by the ScreenCell device. Black round dots correspond to 8 μm pores of the filter (D). Scale bars A, B, C, D = 20 μm.</p