源自乳腺癌患者外周血的循环肿瘤细胞系的建立与鉴定
Establishment and Characterization of a CTC Cell Line from Peripheral Blood of Breast Cancer Patient.
作者信息
Zhao Pan, Zhou Wenbin, Liu Chang, Zhang Huirong, Cheng Zhiqiang, Wu Weiqing, Liu Kaisheng, Hu Hong, Zhong Caineng, Zhang Yayuan, Zhou Dongxian, Liu Feiyuan, Dai Yong, Wang Jianhong, Zou Chang
机构信息
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.
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.
出版信息
J Cancer. 2019 Oct 15;10(24):6095-6104. doi: 10.7150/jca.33157. eCollection 2019.
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 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 and 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.
背景
基于循环肿瘤细胞（CTC）的患者来源细胞是研究癌症分子基础的理想模型。然而，CTC的稀有性和异质性以及原代培养的困难限制了它们的实际应用。建立高效的培养方法并对CTC进行功能表征对于癌症研究至关重要。为此，我们开发了一种用于分离、扩增和鉴定乳腺癌CTC的实验方案。
方法
从一名乳腺癌患者的外周血细胞中建立了CTC-3细胞系。进行了核型分析。通过流式细胞术、定量实时PCR和蛋白质印迹评估分子特征。通过细胞生长和肿瘤球形成试验以及小鼠异种移植模型评估CTC-3细胞形成的肿瘤的特征。通过免疫组织化学、免疫荧光分析和苏木精和伊红染色对肿瘤进行分析。
结果
与广泛使用的MCF-7乳腺癌细胞系相比，CTC-3细胞系在体外和体内均表现出更具侵袭性的生长。CTC-3细胞对化疗药物也更具抗性，基因谱分析表明与MCF-7细胞相比，上皮-间质转化和干性标志物的表达水平更高。
结论
与现有细胞系相比，CTC-3细胞是研究乳腺癌恶性行为的更好模型。
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