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Review Open Access

Advances in isolation and detection technologies and immunotherapy applications of circulating tumor cells (Review)

  • Authors:
    • Ping Zhou
    • Qin Ye
    • Yiyun Huang
    • Yumei Feng
    • Li Zhou
    • Ke Xie
  • View Affiliations / Copyright

    Affiliations: Department of Oncology, Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, Sichuan 610054, P.R. China, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, P.R. China
    Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 57
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    Published online on: January 28, 2026
       https://doi.org/10.3892/or.2026.9062
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Abstract

Circulating tumor cells (CTCs) are shed from the primary tumor into the peripheral bloodstream, where they play crucial roles in tumor metastasis and recurrence. As a cornerstone of liquid biopsy, CTCs hold significant potential for early tumor diagnosis, therapeutic response monitoring, and prognosis. However, the rarity and heterogeneity of CTCs pose considerable challenges for their isolation and enrichment. Additionally, their predictive usefulness in tumor immunotherapy remains relatively limited. The present review summarizes recent advancements in CTC isolation and detection technologies, explores their clinical applications in immunotherapy, and discusses current challenges alongside potential strategies for improvement. The integration of these technologies into clinical practice could pave the way for more personalized and precise cancer treatment strategies in the future.

View Figures

Figure 1

A brief chronological overview of the
development of CTC enrichment techniques. Image was created using
www.Figdraw.com. CTCs, circulating tumor
cells.

Figure 2

Brief process of CTC technology. (A)
Separation techniques: These can be divided into three categories:
Based on physical properties, based on biological properties, and
combining physical properties and biological properties. (B)
Detection techniques: RT-PCR, flow cytometry and NGS. (C)
Downstream analyses: Genomics, proteomics and transcriptomics.
Image was created using www.Figdraw.com. CTCs, circulating tumor cells;
RT-PCR, reverse transcription-polymerase chain reaction; NGS,
next-generation sequencing.

Figure 3

Schematic diagram of the Cell Search
system for detecting CTCs. (A) Blood is drawn into a Cell Save
protection tube containing EDTA and cytoprotectant. (B) A 7.5-ml
aliquot of blood is transferred to a separate tube for
centrifugation. (C) The samples are processed using the
CELLTRACKS®AUTOPREP® system, during which
plasma is aspirated and the cells are resuspended in a buffer
solution. (D) Magnetic nanoparticles coupled with EpCAM antibodies
are added to bind EpCAM-positive cells, enriching for
epithelial-derived CTCs. The bead-bound cells are then separated
from other cells via magnetic force. (E) The CTCs are stained with
antibodies targeting CK8, CK18 and CK19. CD45-positive cells
(leukocytes) are excluded from the analysis. (F) The nuclei are
stained with DAPI for identification. (G) Magnetic separation is
performed to isolate EpCAM-positive cells bound to the magnetic
beads. (H) Cells that are CK-positive, DAPI-positive and
CD45-negative are identified as CTCs and selected for further
analysis. Image was created using www.Figdraw.com. CTCs, circulating tumor cells;
EpCAMs, epithelial cell adhesion molecules.

Figure 4

Mechanisms underlying CTC immune
evasion. (A) Tumor cells are shed from the primary focus. (B)
Dislodged tumor cells traverse the mesenchymal connective tissue,
penetrate the blood vessel walls and enter the circulation as CTCs.
(C) Tumor cells that enter the bloodstream remain in circulation
temporarily before extravasating (exiting the circulatory system)
to attach to other organs or tissues. These cells then begin to
proliferate, eventually forming new metastases. (D) In addition to
individual CTCs, clusters of CTCs exhibit significantly higher
metastatic potential. (E) Immune escape mechanisms: These include
modifications of MHC-I expression; ‘don't eat me’ signaling via
CD47, upregulation of PD-L1 expression, and the expression of FasL,
which induces apoptosis in Fas-positive effector immune cells.
Image was created using www.Figdraw.com. CTCs, circulating tumor cells; PD-1,
programmed cell death protein 1; PD-L1, programmed cell death
ligand 1; MHC, major histocompatibility complex; TCR, T cell
receptor; MDSC, myeloid-derived suppressor cell.
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Copy and paste a formatted citation
Spandidos Publications style
Zhou P, Ye Q, Huang Y, Feng Y, Zhou L and Xie K: <p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>. Oncol Rep 55: 57, 2026.
APA
Zhou, P., Ye, Q., Huang, Y., Feng, Y., Zhou, L., & Xie, K. (2026). <p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>. Oncology Reports, 55, 57. https://doi.org/10.3892/or.2026.9062
MLA
Zhou, P., Ye, Q., Huang, Y., Feng, Y., Zhou, L., Xie, K."<p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>". Oncology Reports 55.4 (2026): 57.
Chicago
Zhou, P., Ye, Q., Huang, Y., Feng, Y., Zhou, L., Xie, K."<p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>". Oncology Reports 55, no. 4 (2026): 57. https://doi.org/10.3892/or.2026.9062
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Spandidos Publications style
Zhou P, Ye Q, Huang Y, Feng Y, Zhou L and Xie K: <p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>. Oncol Rep 55: 57, 2026.
APA
Zhou, P., Ye, Q., Huang, Y., Feng, Y., Zhou, L., & Xie, K. (2026). <p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>. Oncology Reports, 55, 57. https://doi.org/10.3892/or.2026.9062
MLA
Zhou, P., Ye, Q., Huang, Y., Feng, Y., Zhou, L., Xie, K."<p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>". Oncology Reports 55.4 (2026): 57.
Chicago
Zhou, P., Ye, Q., Huang, Y., Feng, Y., Zhou, L., Xie, K."<p>Advances in isolation and detection technologies and immunotherapy applications of circulating&nbsp;tumor cells (Review)</p>". Oncology Reports 55, no. 4 (2026): 57. https://doi.org/10.3892/or.2026.9062
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