Circulating tumor cell isolation Kimchi ET[Author]
Cancer cells can be detected in the blood as circulating tumor cells. Author Eric T Kimchi and others report on their findings in Cancer Biol Ther. 2015 Mar 25:0.
Kaifi JT1, Kunkel M, Das A, Harouaka RA, Dicker DT, Li G, Zhu J, Clawson GA, Yang Z, Reed MF, Gusani NJ, Kimchi ET, Staveley-O'Carroll KF, Zheng SY, El-Deiry WS.
Circulating tumor cell isolation during resection of colorectal cancer lung and liver metastases: a prospective trial with different detection techniques.
ABSTRACT
Background: Colorectal cancer (CRC) metastasectomy improves survival, however most patient develop recurrences. Circulating tumor cells (CTCs) are an independent prognostic marker in stage IV CRC. We hypothesized that CTCs can be enriched during metastasectomy applying different isolation techniques.
Methods: 25 CRC patients undergoing liver (16 (64%)) or lung (9 (36%)) metastasectomy were prospectively enrolled (clinicaltrial.gov identifier: NCT01722903). Central venous (liver) or radial artery (lung) tumor outflow blood (7.5 ml) was collected at incision, during resection, 30 min after resection, and on postoperative day (POD) 1. CTCs were quantified with 1. EpCAM-based CellSearch→ system and 2. size-based isolation with a novel filter device (FMSA). CTCs were immunohistochemically identified using CellSearch→‘s criteria (cytokeratin 8/18/19+, CD45- cells containing a nucleus (DAPI+)). CTCs were also enriched with a centrifugation technique (OncoQuick→).
Results: CTC numbers peaked during the resection with the FMSA in contrast to CellSearch→ (mean CTC number during resection: FMSA: 22.56 (SEM 7.48) (p=0.0281), CellSearch→: 0.87 (SEM ±0.44) (p=0.3018)). Comparing the two techniques, CTC quantity was significantly higher with the FMSA device (range 0-101) than CellSearch→ (range 0-9) at each of the four time points examined (p<0.05). Immunofluorescence staining of cultured CTCs revealed that CTCs have a combined epithelial (CK8/18/19) and macrophage (CD45/CD14) phenotype.
Conclusions: Blood sampling during CRC metastasis resection is an opportunity to increase CTC capture efficiency. CTC isolation with the FMSA yields more CTCs than the CellSearch→ system. Future studies should focus on characterization of single CTCs to identify targets for molecular therapy and immune escape mechanisms of cancer cells.
Kaifi JT1, Kunkel M, Das A, Harouaka RA, Dicker DT, Li G, Zhu J, Clawson GA, Yang Z, Reed MF, Gusani NJ, Kimchi ET, Staveley-O'Carroll KF, Zheng SY, El-Deiry WS.
Circulating tumor cell isolation during resection of colorectal cancer lung and liver metastases: a prospective trial with different detection techniques.
ABSTRACT
Background: Colorectal cancer (CRC) metastasectomy improves survival, however most patient develop recurrences. Circulating tumor cells (CTCs) are an independent prognostic marker in stage IV CRC. We hypothesized that CTCs can be enriched during metastasectomy applying different isolation techniques.
Methods: 25 CRC patients undergoing liver (16 (64%)) or lung (9 (36%)) metastasectomy were prospectively enrolled (clinicaltrial.gov identifier: NCT01722903). Central venous (liver) or radial artery (lung) tumor outflow blood (7.5 ml) was collected at incision, during resection, 30 min after resection, and on postoperative day (POD) 1. CTCs were quantified with 1. EpCAM-based CellSearch→ system and 2. size-based isolation with a novel filter device (FMSA). CTCs were immunohistochemically identified using CellSearch→‘s criteria (cytokeratin 8/18/19+, CD45- cells containing a nucleus (DAPI+)). CTCs were also enriched with a centrifugation technique (OncoQuick→).
Results: CTC numbers peaked during the resection with the FMSA in contrast to CellSearch→ (mean CTC number during resection: FMSA: 22.56 (SEM 7.48) (p=0.0281), CellSearch→: 0.87 (SEM ±0.44) (p=0.3018)). Comparing the two techniques, CTC quantity was significantly higher with the FMSA device (range 0-101) than CellSearch→ (range 0-9) at each of the four time points examined (p<0.05). Immunofluorescence staining of cultured CTCs revealed that CTCs have a combined epithelial (CK8/18/19) and macrophage (CD45/CD14) phenotype.
Conclusions: Blood sampling during CRC metastasis resection is an opportunity to increase CTC capture efficiency. CTC isolation with the FMSA yields more CTCs than the CellSearch→ system. Future studies should focus on characterization of single CTCs to identify targets for molecular therapy and immune escape mechanisms of cancer cells.
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