Lee Soo Jeong, Lee Cham Han, Choi Sung Ho, Ahn Sei Hyun, Son Byung Ho, Lee Jong Won, Yu Jong Han, Kwon Nak-Jung, Lee Woo Chung, Yang Kap-Seok, Lee Dong Hyoung, Han Du Yeol, Choi Mi So, Park Pyeong-Soo, Lee Hyun Kyung, Kim Myoung Shin, Lee Jinseon, Jeon Byung Hee
Cytogen, Inc., Songpa-gu, Seoul 138-961, Republic of Korea.
Department of Surgery, Asan Medical Center Affiliated to The University of Uslan College of Medicine, Songpa-gu, Seoul 138-736, Republic of Korea.
Oncol Lett. 2017 May;13(5):3025-3031. doi: 10.3892/ol.2017.5807. Epub 2017 Mar 6.
Liquid biopsy isolation of circulating tumor cells (CTCs) allows the genomic analysis of CTCs, which is useful in the determination of personalized cancer therapy. In the present study, CTCs from patients with breast cancer were enriched and successfully analyzed using cancer gene panel analysis. Blood samples from 11 patients with breast cancer were collected and CTCs enriched for using size-based filtration. The enriched CTCs were analyzed using immunofluorescence staining with antibodies directed against epithelial cell adhesion molecule (EpCAM) and cluster of differentiation 45. The genomic DNA of CTCs was extracted, amplified and 50 genes screened for mutations using the Ion AmpliSeq™ Cancer Hotspot Panel v2. EpCAM staining detected CTCs in 10/11 patients and the average CTC count was 3.9 in 5 ml blood. The average purity of enriched CTCs was 14.2±29.4% and the average amount of amplified DNA was 28.6±11.9 µg. Catalogue Of Somatic Mutations In Cancer mutations were detected in the CTCs and included IDH2, TP53, NRAS, IDH1, PDGFRA, HRAS, STK11, EGFR, PTEN, MLH1, PIK3CA, CDKN2A, KIT and SMARCB1. In conclusion, a novel size-based filtration approach for the isolation of CTCs was evaluated and successfully applied for the genomic analysis of CTCs from patients with breast cancer.
循环肿瘤细胞（CTC）的液体活检分离技术可实现对CTC的基因组分析，这有助于确定个性化癌症治疗方案。在本研究中，采用癌症基因panel分析对乳腺癌患者的CTC进行了富集和成功分析。收集了11例乳腺癌患者的血样，通过基于尺寸的过滤方法富集CTC。使用针对上皮细胞粘附分子（EpCAM）和分化簇45的抗体进行免疫荧光染色，对富集的CTC进行分析。提取、扩增CTC的基因组DNA，并使用Ion AmpliSeq™癌症热点panel v2筛选50个基因的突变情况。EpCAM染色在11例患者中的10例中检测到了CTC，5ml血液中CTC的平均计数为3.9。富集的CTC的平均纯度为14.2±29.4%，扩增DNA的平均量为28.6±11.9μg。在CTC中检测到了癌症体细胞突变目录中的突变，包括IDH2、TP53、NRAS、IDH1、PDGFRA、HRAS、STK11、EGFR、PTEN、MLH1、PIK3CA、CDKN2A、KIT和SMARCB1。总之，评估了一种用于分离CTC的新型基于尺寸的过滤方法，并成功将其应用于乳腺癌患者CTC的基因组分析。
