Ethical statements
Cell culture
Mouse experiments
Metaphase preparation and spectral karyotyping analysis
Flow cytometry
Immunocytochemistry, IHC, and EMT scoring
Proliferation assay, MTT assay, and 2D and 3D colony formation assays
Adhesion, invasion, and scratch assay
Generation of single cell–derived clones
Quantitative real-time polymerase chain reaction
Patients
SE of CTCs and DTCs
Identification of CTCs and DTCs by iFISH
Collection of single CTC and amplification
Quality control of WGA products and library preparation
Whole-genome sequencing, CNV, and GO analysis
Statistical analysis
The objectives of the present study were to assess the association of differing EMT phenotypes in systemic cancer cells of MBC with their ability to form lung metastases in vitro, ex vivo, and in vivo in the 4T1 MBC syngeneic mouse model. Our major focus was on CTCs and DTCs, their EMT phenotypes, proliferation, adhesion, migration, invasion, tumorigenesis, and metastatic potential. In addition, the objective of the study was to analyze EMT phenotypes of systemic CTCs and DTCs from primary patients with MBC and to correlate EMT phenotypes with the metastatic status and clinical outcome of the patients.