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Circulating tumor cells (CTCs) are tumor cells shed from either primary tumors or its metastases that circulate in the peripheral blood of patients with metastatic cancers. The molecular characterization of the CTCs is critical to identifying the key drivers of cancer metastasis and devising therapeutic approaches. However, the molecular characterization of CTCs is difficult to achieve because their isolation is a major technological challenge.    Methods  CTCs from two triple negative breast cancer patients were enriched using CellSearch and single cells selected by DEPArray™. A  TP53  R110 fs*13 mutation identified by next generation sequencing in the breast and chest skin biopsies of both patients was studied in single CTCs.    Results  From 6 single CTC isolated from one patient, 1 CTC had  TP53  R110 delC, 1 CTC showed the  TP53  R110 delG mutation, and the remaining 4 single CTCs showed the wild type p53 sequence; a pool of 14 CTCs isolated from the same patient also showed  TP53  R110 delC mutation. In the tumor breast tissue of this patient, only the  TP53  R110 delG mutation was detected. In the second patient a  TP53  R110 delC mutation was detected in the chest wall skin biopsy; from the peripheral blood of this patient, 5 single CTC and 6 clusters of 2 to 6 CTCs were isolated; 3 of the 5 single CTCs showed the  TP53  R110 delC mutation and 2 CTCs showed the wild type  TP53  allele; from the clusters, 5 showed the  TP53  R110 delC mutation, and 1 cluster the wild type  TP53  allele. Single white blood cells isolated as controls from both patients only showed the wild type  TP53  allele.    Conclusions  We are able to isolate uncontaminated CTCs and achieve single cell molecular analysis. Our studies showed the presence of different CTC sub-clones in patients with metastatic breast cancer. Some CTCs had the same TP53 mutation as their matching tumor samples although others showed either a different TP53 mutation or the wild type allele. Our results indicate that CTCs could represent a non-invasive source of cancer cells from which to determine genetic markers of the disease progression and potential therapeutic targets.    Electronic supplementary material  The online version of this article (doi:10.1186/s13058-014-0445-3) contains supplementary material, which is available to authorized users.

TP53 mutations detected in circulating tumor cells present in the blood of metastatic triple negative breast cancer patients