Isolation and Characterization of Circulating XXXX Cells in Squamous Cell Carcinoma of the Lung Using a Non- @entity5960 -Based Capture Method.

@entity1 @entity5960 @entity5 @entity1544 @entity2277 @entity7167
INTRODUCTION: The exclusion of circulating @entity5 cells (CTCs) that have lost epithelial antigens during the epithelial-to-mesenchymal transition (EMT) process by using @entity5960 ( @entity5960 ) based capture methods is still a matter of debate. In this study, cells obtained after depletion procedure from blood samples of @entity1544 ( @entity1544 ) @entity1 were identified based on morphology and characterized with the combination of FISH assessment and immunophenotypic profile. MATERIALS AND METHODS: Five mL blood samples, collected from 55 advanced @entity1544 @entity1 , were analyzed by a non- @entity5960 -based capture method. After depletion of leukocytes and erythroid cells, the negative fraction was characterized by both FISH using a @entity7167 ( @entity7167 ) probe and by immunocytochemistry. Thirty healthy donors were also tested. RESULTS: Based on morphology (nuclear dimension >= 10  m, shape and hypercromatic aspect) suspicious circulating cells clearly distinguishable from contaminant leukocytes were observed in 49/55 (89%) @entity1544 @entity1 . Thirty-four of the 44 (77%) samples evaluable for @entity7167 FISH showed >=  6 @entity7167 gene copy number on average per cell. @entity2277 expression involved 43% (18/42) of pooled circulating @entity1544 cells, whereas only 29% (14/48) were @entity5960 positive. Confocal microscopy confirmed the localization of @entity7167 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 @entity1544 @entity1 was found. CONCLUSIONS: 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 @entity5 cells in advanced @entity1544 . The finding of cells with neither @entity5960 or EMT phenotype, retrieved after non- @entity5960 -based systems, underlines the presence of suspicious elements in the blood of both @entity1544 @entity1 and healthy donors. Further phenotyping and molecular analyses are necessary to fully characterize these circulating elements.
