Τίτλος:
Direct comparison of size-dependent versus EpCAM-dependent CTC enrichment at the gene expression and DNA methylation level in head and neck squamous cell carcinoma
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
We directly compared two different approaches used for Circulating Tumor Cell (CTC) isolation, a size-dependent microfluidic system versus an EpCAM-dependent positive selection for downstream molecular characterization of CTC both at the gene expression and DNA methylation level in Head and Neck Squamous Cell Carcinoma (HNSCC). A size-dependent microfluidic device (Parsortix, ANGLE) and an EpCAM-dependent positive immune-magnetic isolation procedure were applied in parallel, using 10 mL PB from 50 HNSCC patients and 18 healthy donors. Total RNA was isolated from enriched CTCs and RT-qPCR was used to study the expression levels of CK-19, PD-L1, EGFR, TWIST1, CDH2 and B2M (reference gene). Real time methylation specific PCR (MSP) was used to study the methylation status of RASSF1A and MLL3 genes. In identical blood draws, the label-free size-dependent CTC-isolation system was superior in terms of sensitivity when compared to the EpCAM-dependent CTC enrichment, since a significantly higher percentage of identical PB samples was found positive at the gene expression and DNA methylation level, while the specificity was not affected. Our results indicate that future studies focused on the evaluation of clinical utility of CTC molecular characterization in HNSCC should be based on size-dependent enrichment approaches. © 2020, The Author(s).
Συγγραφείς:
Zavridou, M.
Mastoraki, S.
Strati, A.
Koutsodontis, G.
Klinakis, A.
Psyrri, A.
Lianidou, E.
Περιοδικό:
Scientific Reports
Εκδότης:
Institute of Geographic Sciences and Natural Resources Research
Λέξεις-κλειδιά:
epithelial cell adhesion molecule; tumor marker, cell size; comparative study; DNA methylation; gene expression regulation; genetics; human; metabolism; pathology; tumor cell line; tumor embolism, Biomarkers, Tumor; Cell Line, Tumor; Cell Size; DNA Methylation; Epithelial Cell Adhesion Molecule; Gene Expression Regulation, Neoplastic; Humans; Neoplastic Cells, Circulating; Squamous Cell Carcinoma of Head and Neck
DOI:
10.1038/s41598-020-63055-y
