A liquid biopsy-based method for the detection and quantification of circulating tumor cells in surgical osteosarcoma patients

Zhang,Haoqiang; Gao,Peng; Xiao,Xin; Heger,Michal; Geng,Lei; Fan,Bo; Yuan,Yulin; Huang,Chen; Chen,Guojing; Liu,Yao; Hu,Yongchen; Yu,Xiuchun; Wu,Sujia; Wang,Ling; Wang,Zhen

doi:10.3892/ijo.2017.3905

A liquid biopsy-based method for the detection and quantification of circulating tumor cells in surgical osteosarcoma patients

Authors:
- Haoqiang Zhang
- Peng Gao
- Xin Xiao
- Michal Heger
- Lei Geng
- Bo Fan
- Yulin Yuan
- Chen Huang
- Guojing Chen
- Yao Liu
- Yongchen Hu
- Xiuchun Yu
- Sujia Wu
- Ling Wang
- Zhen Wang
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Affiliations: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Experimental Surgery, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands, Membrane Biochemistry and Biophysics, Institute of Biomembranes, Utrecht University, 3584CH Utrecht, The Netherlands, Department of Orthopedic Oncology, Tianjin Hospital, Tianjin 300210, P.R. China, Department of Orthopedics, The General Hospital of Jinan Military Commanding Region, Jinan, Shandong 250031, P.R. China, Department of Orthopedics, The General Hospital of Nanjing Military Commanding Region, Nanjing, Jiangsu 210008, P.R. China, Department of Vascular and Endocrine Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China
Published online on: March 8, 2017 https://doi.org/10.3892/ijo.2017.3905
Pages: 1075-1086
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A method for the enumeration and quantification of osteosarcoma (OS) circulating tumor cells (CTCs) is currently not available. A correlation between the number of CTCs and progression-free survival (PFS) has been established for other cancers, but not for OS CTCs. A method was therefore developed for CTC quantification in OS and validated in a prospective cohort of surgical patients with primary and recurrent/metastatic OS (N=23). Human OS cells, acting as CTCs, were enumerated from spiked human peripheral blood (PB) following erythrocyte and leukocyte depletion. The OS cells were quantified microscopically based on aneuploidy and a CK18-/CD45- phenotype. Aneuploidy was assayed by fluorescence in situ hybridization (FISH) using fluorescence-labeled alpha-satellite probes for the centromeres of chromosome (CEP 8). CK18 and CD45 phenotyping was performed with immunocytochemistry. HOS cells in spiked PB could be effectively retrieved with the FISH-based enumeration method, which was subsequently employed in an OS patient cohort. PB of recurrent/metastatic OS patients contained more CTCs than the PB of primary OS patients. OS patients with ≥2 CTCs per 7.5 ml of PB had worse PFS than patients whose PB contained <2 CTCs. In 2 cases, CTCs were present in PB of OS patients with negative X-ray and chest CT scans. In conclusion, our method was able to quantitate CTCs in liquid biopsies of OS patients. The number of CTCs has diagnostic and prognostic value.

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