Abstract

Circulating tumor cells (CTCs) are of recognized importance for diagnosis and prognosis of cancer patients. With melanoma, most studies do not show any clear relationship between CTC levels and stage of disease. Here, CTCs were enriched (~400X) from blood of melanoma patients using a simple centrifugation device (OncoQuick), and 4 melanocyte target RNAs (TYR, MLANA, MITF, and MIF) were quantified using QPCR. Approximately one-third of melanoma patients had elevated MIF and MLANA transcripts (p<0.0001 and p<0.001, respectively) compared with healthy controls. In contrast, healthy controls had uniformly higher levels of TYR and MITF than melanoma patients (p<0.0001). There was a marked shift of leukocytes into the CTC-enriched fractions (a 430% increase in RNA recovery, p<0.001), and no relationship between CTC levels and stage of disease was found. CTCs were captured on microfabricated filters and cultured. Captured melanoma CTCs were large cells, and consisted of 2 subpopulations, based on immunoreactivity. One subpopulation (~50%) stained for both pan-cytokeratin (KRT) markers and the common leukocyte marker CD-45, whereas the second subpopulation stained for only KRT. Since similar cells are described in many cancers, we also examined blood from colorectal and pancreatic cancer patients. We observed analogous results, with most captured CTCs staining for both CD-45/KRT markers (and for the monocyte differentiation marker CD-14). Our results suggest that immature melanocyte-related cells (expressing TYR and MITF RNA) may circulate in healthy controls, although they are not readily detectable without considerable enrichment. Further, as early-stage melanomas develop, immature melanocyte migration into the blood is somehow curtailed, whereas a significant proportion of patients develop elevated CTC levels (based on MIF and MLANA RNAs). The nature of the captured CTCs is consistent with literature describing leukocyte/macrophage-tumor cell fusion hybrids, and their role in metastatic progression.

Original languageEnglish (US)
Article numbere41052
JournalPloS one
Volume7
Issue number7
DOIs
StatePublished - Jul 19 2012

Fingerprint

Circulating Neoplastic Cells
melanoma
Tumors
Melanoma
Cells
Melanocytes
melanocytes
RNA
Leukocytes
leukocytes
Blood
blood
neoplasm cells
immatures
Neoplasms
pancreatic neoplasms
Cell Fusion
cell fusion
Differentiation Antigens
neoplasms

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Clawson, G., Kimchi, E., Patrick, S. D., Xin, P., Harouaka, R., Zheng, S., ... Thiboutot, D. (2012). Circulating tumor cells in melanoma patients. PloS one, 7(7), [e41052]. https://doi.org/10.1371/journal.pone.0041052
Clawson, Gary ; Kimchi, Eric ; Patrick, Susan D. ; Xin, Ping ; Harouaka, Ramdane ; Zheng, Siyang ; Berg, Arthur ; Schell, Todd ; Staveley-O'Carroll, Kevin F. ; Neves, Rogerio ; Mosca, Paul J. ; Thiboutot, Diane. / Circulating tumor cells in melanoma patients. In: PloS one. 2012 ; Vol. 7, No. 7.
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title = "Circulating tumor cells in melanoma patients",
abstract = "Circulating tumor cells (CTCs) are of recognized importance for diagnosis and prognosis of cancer patients. With melanoma, most studies do not show any clear relationship between CTC levels and stage of disease. Here, CTCs were enriched (~400X) from blood of melanoma patients using a simple centrifugation device (OncoQuick), and 4 melanocyte target RNAs (TYR, MLANA, MITF, and MIF) were quantified using QPCR. Approximately one-third of melanoma patients had elevated MIF and MLANA transcripts (p<0.0001 and p<0.001, respectively) compared with healthy controls. In contrast, healthy controls had uniformly higher levels of TYR and MITF than melanoma patients (p<0.0001). There was a marked shift of leukocytes into the CTC-enriched fractions (a 430{\%} increase in RNA recovery, p<0.001), and no relationship between CTC levels and stage of disease was found. CTCs were captured on microfabricated filters and cultured. Captured melanoma CTCs were large cells, and consisted of 2 subpopulations, based on immunoreactivity. One subpopulation (~50{\%}) stained for both pan-cytokeratin (KRT) markers and the common leukocyte marker CD-45, whereas the second subpopulation stained for only KRT. Since similar cells are described in many cancers, we also examined blood from colorectal and pancreatic cancer patients. We observed analogous results, with most captured CTCs staining for both CD-45/KRT markers (and for the monocyte differentiation marker CD-14). Our results suggest that immature melanocyte-related cells (expressing TYR and MITF RNA) may circulate in healthy controls, although they are not readily detectable without considerable enrichment. Further, as early-stage melanomas develop, immature melanocyte migration into the blood is somehow curtailed, whereas a significant proportion of patients develop elevated CTC levels (based on MIF and MLANA RNAs). The nature of the captured CTCs is consistent with literature describing leukocyte/macrophage-tumor cell fusion hybrids, and their role in metastatic progression.",
author = "Gary Clawson and Eric Kimchi and Patrick, {Susan D.} and Ping Xin and Ramdane Harouaka and Siyang Zheng and Arthur Berg and Todd Schell and Staveley-O'Carroll, {Kevin F.} and Rogerio Neves and Mosca, {Paul J.} and Diane Thiboutot",
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Clawson, G, Kimchi, E, Patrick, SD, Xin, P, Harouaka, R, Zheng, S, Berg, A, Schell, T, Staveley-O'Carroll, KF, Neves, R, Mosca, PJ & Thiboutot, D 2012, 'Circulating tumor cells in melanoma patients', PloS one, vol. 7, no. 7, e41052. https://doi.org/10.1371/journal.pone.0041052

Circulating tumor cells in melanoma patients. / Clawson, Gary; Kimchi, Eric; Patrick, Susan D.; Xin, Ping; Harouaka, Ramdane; Zheng, Siyang; Berg, Arthur; Schell, Todd; Staveley-O'Carroll, Kevin F.; Neves, Rogerio; Mosca, Paul J.; Thiboutot, Diane.

In: PloS one, Vol. 7, No. 7, e41052, 19.07.2012.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Circulating tumor cells in melanoma patients

AU - Clawson, Gary

AU - Kimchi, Eric

AU - Patrick, Susan D.

AU - Xin, Ping

AU - Harouaka, Ramdane

AU - Zheng, Siyang

AU - Berg, Arthur

AU - Schell, Todd

AU - Staveley-O'Carroll, Kevin F.

AU - Neves, Rogerio

AU - Mosca, Paul J.

AU - Thiboutot, Diane

PY - 2012/7/19

Y1 - 2012/7/19

N2 - Circulating tumor cells (CTCs) are of recognized importance for diagnosis and prognosis of cancer patients. With melanoma, most studies do not show any clear relationship between CTC levels and stage of disease. Here, CTCs were enriched (~400X) from blood of melanoma patients using a simple centrifugation device (OncoQuick), and 4 melanocyte target RNAs (TYR, MLANA, MITF, and MIF) were quantified using QPCR. Approximately one-third of melanoma patients had elevated MIF and MLANA transcripts (p<0.0001 and p<0.001, respectively) compared with healthy controls. In contrast, healthy controls had uniformly higher levels of TYR and MITF than melanoma patients (p<0.0001). There was a marked shift of leukocytes into the CTC-enriched fractions (a 430% increase in RNA recovery, p<0.001), and no relationship between CTC levels and stage of disease was found. CTCs were captured on microfabricated filters and cultured. Captured melanoma CTCs were large cells, and consisted of 2 subpopulations, based on immunoreactivity. One subpopulation (~50%) stained for both pan-cytokeratin (KRT) markers and the common leukocyte marker CD-45, whereas the second subpopulation stained for only KRT. Since similar cells are described in many cancers, we also examined blood from colorectal and pancreatic cancer patients. We observed analogous results, with most captured CTCs staining for both CD-45/KRT markers (and for the monocyte differentiation marker CD-14). Our results suggest that immature melanocyte-related cells (expressing TYR and MITF RNA) may circulate in healthy controls, although they are not readily detectable without considerable enrichment. Further, as early-stage melanomas develop, immature melanocyte migration into the blood is somehow curtailed, whereas a significant proportion of patients develop elevated CTC levels (based on MIF and MLANA RNAs). The nature of the captured CTCs is consistent with literature describing leukocyte/macrophage-tumor cell fusion hybrids, and their role in metastatic progression.

AB - Circulating tumor cells (CTCs) are of recognized importance for diagnosis and prognosis of cancer patients. With melanoma, most studies do not show any clear relationship between CTC levels and stage of disease. Here, CTCs were enriched (~400X) from blood of melanoma patients using a simple centrifugation device (OncoQuick), and 4 melanocyte target RNAs (TYR, MLANA, MITF, and MIF) were quantified using QPCR. Approximately one-third of melanoma patients had elevated MIF and MLANA transcripts (p<0.0001 and p<0.001, respectively) compared with healthy controls. In contrast, healthy controls had uniformly higher levels of TYR and MITF than melanoma patients (p<0.0001). There was a marked shift of leukocytes into the CTC-enriched fractions (a 430% increase in RNA recovery, p<0.001), and no relationship between CTC levels and stage of disease was found. CTCs were captured on microfabricated filters and cultured. Captured melanoma CTCs were large cells, and consisted of 2 subpopulations, based on immunoreactivity. One subpopulation (~50%) stained for both pan-cytokeratin (KRT) markers and the common leukocyte marker CD-45, whereas the second subpopulation stained for only KRT. Since similar cells are described in many cancers, we also examined blood from colorectal and pancreatic cancer patients. We observed analogous results, with most captured CTCs staining for both CD-45/KRT markers (and for the monocyte differentiation marker CD-14). Our results suggest that immature melanocyte-related cells (expressing TYR and MITF RNA) may circulate in healthy controls, although they are not readily detectable without considerable enrichment. Further, as early-stage melanomas develop, immature melanocyte migration into the blood is somehow curtailed, whereas a significant proportion of patients develop elevated CTC levels (based on MIF and MLANA RNAs). The nature of the captured CTCs is consistent with literature describing leukocyte/macrophage-tumor cell fusion hybrids, and their role in metastatic progression.

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Clawson G, Kimchi E, Patrick SD, Xin P, Harouaka R, Zheng S et al. Circulating tumor cells in melanoma patients. PloS one. 2012 Jul 19;7(7). e41052. https://doi.org/10.1371/journal.pone.0041052