Background. Non-small cell lung cancer (NSCLC) is the most common cause of cancer-related mortality; nevertheless, there are few\ndata regarding detection of circulating tumor cells (CTCs) in NSCLC, compared to other kinds of cancers in which their prognostic\nroles have already been defined. This difference is likely due to detection methods based on the epithelial marker expression which\nignore CTCs undergoing epithelial-mesenchymal transition (CTCsEMT). Methods. After optimization of the test with spiking\nexperiments of A549 cells undergoing TGF-�²1-induced EMT (A549EMT), the CTCsEMT were enriched by immunomagnetic\ndepletion of leukocytes and then characterized by a RT-PCR assay based on the retrieval of epithelial and EMT-related genes.\nBlood samples from ten metastatic NSCLC patients before starting treatment and during chemotherapy were used to test this\napproach by longitudinal monitoring. Ten age- and sex-matched healthy subjects were also enrolled as controls. Results.\nRecovery experiments of spiked A549EMT cells showed that the RT-PCR assay is a reliable method for detection of CTCsEMT.\nCTCsEMT were detected in three patients at baseline and in six patients after four cycles of cysplatin-based chemotherapy.\nLongitudinal monitoring of three patients showed that the CTCsEMT detection is related to poor therapeutic response.\nConclusions. The RT-PCR-based approach for the evaluation of CTCsEMT phenotype could be a promising and inexpensive tool\nto predict the prognosis and the therapeutic response in NSCLC patients.
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