J Cancer 2019; 10(24):6095-6104. doi:10.7150/jca.33157

Research Paper

Establishment and Characterization of a CTC Cell Line from Peripheral Blood of Breast Cancer Patient

Pan Zhao1*, Wenbin Zhou2*, Chang Liu1,3*, Huirong Zhang1, Zhiqiang Cheng4,5, Weiqing Wu6, Kaisheng Liu1, Hong Hu2, Caineng Zhong2, Yayuan Zhang2, Dongxian Zhou2, Feiyuan Liu1, Yong Dai1✉, Jianhong Wang1,5✉, Chang Zou1,5✉

1. Clinical Medical Research Center, the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University, Shenzhen People's Hospital, Shenzhen 518020, China
2. Department of breast and thyroid surgery, the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University, Shenzhen People's Hospital, Shenzhen 518020, China
3. Central Laboratory, Dalian Municipal Central Hospital, Dalian 116033, China
4. Department of Pathology, the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University, Shenzhen People's Hospital, Shenzhen 518020, China
5. Shenzhen Public Service Platform on Tumor Precision Medicine and Molecular Diagnosis, the Second Clinical Medical College of Jinan University, Shenzhen People's Hospital, Shenzhen 518020, China
6. Department of Health Management, the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University, Shenzhen People's Hospital, Shenzhen 518020, China
* These authors contribute equally to this work

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Citation:
Zhao P, Zhou W, Liu C, Zhang H, Cheng Z, Wu W, Liu K, Hu H, Zhong C, Zhang Y, Zhou D, Liu F, Dai Y, Wang J, Zou C. Establishment and Characterization of a CTC Cell Line from Peripheral Blood of Breast Cancer Patient. J Cancer 2019; 10(24):6095-6104. doi:10.7150/jca.33157. Available from http://www.jcancer.org/v10p6095.htm

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Abstract

Background: Circulating tumor cell (CTC)-based patient-derived cells are ideal models for investigating the molecular basis of cancer. However, the rarity and heterogeneity of CTCs as well as the difficulties of primary culture limit their practical application. Establishing efficient in vitro culture methods and functionally characterizing CTCs is essential for cancer studies. To this end, we developed an experimental protocol for the isolation, expansion, and identification of breast cancer CTCs.

Methods: The CTC-3 cell line was established from peripheral blood cells of a breast cancer patient. A karyotype analysis was performed. The molecular profile was assessed by flow cytometry, quantitative real-time PCR, and western blot. The characteristics of tumors formed by CTC-3 cells were evaluated by cell growth and tumor sphere formation assays and in a mouse xenograft model. The tumors were analyzed by immunohistochemistry, immunofluorescence analysis, and hematoxylin and eosin staining.

Results: The CTC-3 cell line showed more aggressive growth both in vitro and in vivo than the widely used MCF-7 breast cancer cell line. CTC-3 cells were also more resistant to chemotherapeutic agents, and gene profiling indicated higher expression levels of the epithelial-to-mesenchymal transition and stemness markers as compared to MCF-7 cells.

Conclusions: CTC-3 cells are a better model for investigating the malignant behavior of breast cancer than existing cell lines.

Keywords: Circulating tumor cell, Breast cancer, Cancer stem cell, Primary culture