Circulating tumor cells detection in neuroblastoma patients by EpCAM-independent enrichment and immunostaining-fluorescence in situ hybridization

• Published in: EBioMedicine 2018-09, Vol.35, p.244-250
• Main Authors: Liu, Xiangqi, Zhang, Zhenzhen, Zhang, Binbin, Zheng, Yijie, Zheng, Chao, Liu, Baihui, Zheng, Shan, Dong, Kuiran, Dong, Rui
• Format: Article
• Language: English
• Subjects: Child; Child, Preschool; Chromosomes, Human, Pair 8 - genetics; Circulating tumor cells; Epithelial Cell Adhesion Molecule - metabolism; Female; Fluorescent Antibody Technique; Gene Amplification; Humans; In Situ Hybridization, Fluorescence; Logistic Models; Male; Metastasis; Multivariate Analysis; N-Myc Proto-Oncogene Protein - genetics; Neoplasm Metastasis; Neoplastic Cells, Circulating - pathology; Neuroblastoma; Neuroblastoma - genetics; Neuroblastoma - pathology; Neuroblastoma - urine; Phosphopyruvate Hydratase - metabolism; Prognosis; Proportional Hazards Models; Research paper; Risk Factors; Vanilmandelic Acid - urine
• ISSN: 2352-3964; 2352-3964
• DOI: 10.1016/j.ebiom.2018.08.005

Detecting circulating tumor cells (CTCs) has proven valuable for evaluating the prognosis of cancer patients and for studying the mechanisms of treatment resistance. Owing to the lack of universal and specific tumor markers for neuroblastoma (NB), in this prospective study, we adopted an EpCAM-independent method to detect CTCs in the peripheral blood of NB patients. We used an EpCAM-independent assay to delete leukocytes and to enrich the CTCs. CTCs were identified by immunostaining of CD45, DAPI and DNA fluorescence in situ hybridization (FISH) of the centromere of chromosome 8 probe (CEP8). Cells that were DAPI+/CD45-/CEP8 ≥ 3 were considered CTCs. We collected peripheral blood from 28 NB patients as well as clinical and follow-up data. The number of CTCs among the different risk groups were significantly different (p = .0208, Kruskal–Wallis test). Patients with metastasis had more CTCs than those without metastasis (p