Enhanced detection and comprehensive in situ phenotypic characterization of circulating and disseminated heteroploid epithelial and glioma tumor cells
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Feng Ge1,*, Haishi Zhang2,*, Daisy Dandan Wang3, Linda Li3, Peter Ping Lin3
1Department of Thoracic Surgery, Capital Medical University School of Oncology and Chaoyang Hospital, Beijing, China
2Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China
3Cytelligen, San Diego, California, USA
*These authors have contributed equally to this study
Peter Ping Lin, e-mail: [email protected]
Keywords: CTC and DTC subtypes, iFISH, subtraction enrichment, cytokeratin (CK) 18, in situ phenotyping and karyotyping
Received: June 29, 2015 Accepted: July 17, 2015 Published: July 29, 2015
Conventional strategy of anti-EpCAM capture and immunostaining of cytokeratins (CKs) to detect circulating tumor cells (CTCs) is limited by highly heterogeneous and dynamic expression or absence of EpCAM and/or CKs in CTCs. In this study, a novel integrated cellular and molecular approach of subtraction enrichment (SE) and immunostaining-FISH (iFISH) was successfully developed. Both large or small size CTCs and circulating tumor microemboli (CTM) in various biofluid samples including cerebrospinal fluid (CSF) of cancer patients and patient-derived-xenograft (PDX) mouse models were efficiently enriched and comprehensively identified and characterized by SE-iFISH. Non-hematopoietic CTCs with heteroploid chromosome 8 were detected in 87–92% of lung, esophageal and gastric cancer patients. Characterization of CTCs performed by CK18-iFISH showed that CK18, the dual epithelial marker and tumor biomarker, was strong positive in only 14% of lung and 24% of esophageal CTCs, respectively. Unlike conventional methodologies restricted only to the large and/or both EpCAM and CK positive CTCs, SE-iFISH enables efficient enrichment and performing in situ phenotypic and karyotypic identification and characterization of the highly heterogeneous CTC subtypes classified by both chromosome ploidy and the expression of various tumor biomarkers. Each CTC subtype may possess distinct clinical significance relative to tumor metastasis, relapse, therapeutic drug sensitivity or resistance, etc.
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