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Enrichment of circulating melanoma cells (CMCs) using negative selection from patients with metastatic melanoma
- Source :
- Oncotarget, Scopus-Elsevier
- Publication Year :
- 2014
- Publisher :
- Impact Journals LLC, 2014.
-
Abstract
- Circulating tumor cells have emerged as prognostic biomarkers in the treatment of metastatic cancers of epithelial origins viz., breast, colorectal and prostate. These tumors express Epithelial Cell Adhesion Molecule (EpCAM) on their cell surface which is used as an antigen for immunoaffinity capture. However, EpCAM capture technologies are of limited utility for non-epithelial cancers such as melanoma. We report a method to enrich Circulating Melanoma Cells (CMCs) that does not presuppose malignant cell characteristics. CMCs were enriched by centrifugation of blood samples from healthy (N = 10) and patient (N = 11) donors, followed by RBC lysis and immunomagnetic depletion of CD45-positive leukocytes in a specialized magnetic separator. CMCs were identified by immunocytochemistry using Melan-A or S100B as melanoma markers and enumerated using automated microscopy image analyses. Separation was optimized for maximum sensitivity and recovery of CMCs. Our results indicate large number of CMCs in Stage IV melanoma patients. Analysis of survival suggested a trend toward decreased survival with increased number of CMCs. Moreover, melanoma-associated miRs were found to be higher in CMC-enriched fractions in two patients when compared with the unseparated samples, validating this method as applicable for molecular analyses. Negative selection is a promising approach for isolation of CMCs and other EpCAM -negative CTCs, and is amenable to molecular analysis of CMCs. Further studies are required to validate its efficacy at capturing specific circulating cells for genomic analysis, and xenograft studies.
- Subjects :
- Immunocytochemistry
Cell
Cell Separation
S100 Calcium Binding Protein beta Subunit
Biology
Immunomagnetic separation
Real-Time Polymerase Chain Reaction
S100B
Immunoenzyme Techniques
chemistry.chemical_compound
Circulating tumor cell
circulating melanoma cells
MART-1 Antigen
Antigen
medicine
Biomarkers, Tumor
Tumor Cells, Cultured
Humans
RNA, Messenger
magnetic separation
Melanoma
Immunomagnetic Separation
Reverse Transcriptase Polymerase Chain Reaction
Melan-A
negative selection
Epithelial cell adhesion molecule
medicine.disease
Neoplastic Cells, Circulating
Prognosis
Molecular biology
CTC
Survival Rate
MicroRNAs
medicine.anatomical_structure
Oncology
chemistry
Case-Control Studies
Cancer research
Leukocyte Common Antigens
Research Paper
Subjects
Details
- Language :
- English
- ISSN :
- 19492553
- Volume :
- 5
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- Oncotarget
- Accession number :
- edsair.doi.dedup.....346ccf7f4a6506dd5e77fd240a4af3e0