Open Access

Enrichment and mutation detection of circulating tumor cells from blood samples

  • Authors:
    • Ruqin Kou
    • Jian Zhao
    • Priya Gogoi
    • Shannon Carskadon
    • Will Chow
    • Clara Hwang
    • Nallasivam Palanisamy
    • Conrad Leung
    • Yixin Wang
  • View Affiliations

  • Published online on: March 30, 2018     https://doi.org/10.3892/or.2018.6342
  • Pages: 2537-2544
  • Copyright: © Kou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The potential of circulating tumor cells (CTCs) in the diagnosis and prognosis of cancer patients has become increasingly attractive. However, molecular analysis of CTCs is hindered by low sensitivity and a high level of background leukocytes in CTC enrichment technologies. We have developed a novel protocol using a microfluidic device, which enriches and retrieves CTCs from blood samples. The principle of CTC capturing is that tumor cells are larger and less deformable than normal blood cells. To evaluate the potential of utilizing Celsee PREP100 in CTC molecular analysis, we prepared prostate cancer cell lines PC3 and LNCaP, retrieved the captured cells and analyzed them using PCR amplicon sequencing. We were able to recover an average of 79% of 110‑1,100 PC3 and 60‑1,500 LNCaP cells, and detect the p.K139fs*3 deletion of the p53 gene in PC3 cells and p.T877A mutation of the androgen receptor gene in LNCaP cells. Next, we spiked these two types of cells into normal donor blood samples, captured the cells and analyzed them using PCR amplicon sequencing. The PC3 and LNCaP cells were captured and retrieved with the ratio of captured CTCs to the background leukocytes reaching 1:1.5 for PC3 and 1:2.9 for LNCaP cells. We further revealed that the p.K139fs*3 deletion and p.T877A mutation can be detected in the captured PC3 and LNCaP cells, respectively. We successfully validated this approach using clinical blood samples from patients with metastatic prostate cancer. Our results demonstrated a novel approach for CTC enrichment and illustrated the potential of CTC molecular characterization for diagnosis, prognosis and treatment selection of patients with metastatic malignancy.
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June-2018
Volume 39 Issue 6

Print ISSN: 1021-335X
Online ISSN:1791-2431

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Spandidos Publications style
Kou R, Zhao J, Gogoi P, Carskadon S, Chow W, Hwang C, Palanisamy N, Leung C and Wang Y: Enrichment and mutation detection of circulating tumor cells from blood samples. Oncol Rep 39: 2537-2544, 2018.
APA
Kou, R., Zhao, J., Gogoi, P., Carskadon, S., Chow, W., Hwang, C. ... Wang, Y. (2018). Enrichment and mutation detection of circulating tumor cells from blood samples. Oncology Reports, 39, 2537-2544. https://doi.org/10.3892/or.2018.6342
MLA
Kou, R., Zhao, J., Gogoi, P., Carskadon, S., Chow, W., Hwang, C., Palanisamy, N., Leung, C., Wang, Y."Enrichment and mutation detection of circulating tumor cells from blood samples". Oncology Reports 39.6 (2018): 2537-2544.
Chicago
Kou, R., Zhao, J., Gogoi, P., Carskadon, S., Chow, W., Hwang, C., Palanisamy, N., Leung, C., Wang, Y."Enrichment and mutation detection of circulating tumor cells from blood samples". Oncology Reports 39, no. 6 (2018): 2537-2544. https://doi.org/10.3892/or.2018.6342