【文献解读】转移性癌患者的液体活检中循环肿瘤细胞的分析
2017/9/6 创新医学网
文章来源：欧尔意 OAE 微信号
作者：Sofina
原文出处
Potdar PD, Sen K. Profiling of circulating tumor cells in liquid biopsies from metastatic cancer patients. J Cancer Metastasis Treat 2017;3:6-15.
DOI:10.20517/2394-4722.2016.53
摘要
目的:循环肿瘤细胞(CTCs)对肿瘤转移至关重要，对肿瘤患者的临床预后有重要价值。在此，作者试图通过液体活检的方法对CTCs进行枚举和表征。方法:使用Ficoll-Hypaque梯度从转移癌患者的血液样品中分离外周血单核细胞(PBMCs)，并培养以分离和枚举CTCs。培养的CTCs通过光学和相差显微镜进行形态学表征。除了检测其mRNA转移标记的表达外，还检测了Ficoll-Hypaque分离的PBMCs的致瘤性。结果:在培养基中分离出CTCs，并通过在相差显微镜下进行枚举计数，证明了0.01-0.04%的PBMCs是CTCs。CTCs处于休眠状态，有着巨大的卵形、尖刺状的形态。从液体活检中获得的PBMCs显示出贴壁不依赖性生长，在软琼脂试验中形成了大量的菌落。分子分析显示了几种转移基因的表达，但在所有的病人中都没有钙粘蛋白1(编码粘附蛋白)。结论:作者从转移癌患者的液体活检中成功分离、枚举和表征了CTCs。这项研究有助于促进开发液体活检的新的诊断和治疗方法，并应用于转移性癌症中。
关键词：循环肿瘤细胞，液体活检，分子标记，软琼脂试验，转移，转移基因，上皮细胞间质转化， 致瘤的
Aim: Circulating tumor cells (CTCs) are crucial to tumor metastasis and valuable for prediction of clinical outcome in patients with solid tumors. Here, the authors aimed to establish a method for enumeration and characterization of CTCs from liquid biopsies. Methods: Peripheral blood mononuclear cells (PBMCs) were separated from blood samples from patients with metastatic cancer using Ficoll-Hypaque gradients and cultured to isolate and enumerate CTCs. Cultured CTCs were morphologically characterized by light and phase contrast microscopy. The tumorigenicity of Ficoll-Hypaque-separated PBMCs was examined, in addition to their expression of mRNA metastasis markers. Results: CTCs were isolated in culture and enumerated by counting under phase contrast microscopy, demonstrating that 0.01-0.04% of total PBMCs were CTCs. CTCs were dormant, with large, oval-shaped, spiky morphology. PBMCs obtained from liquid biopsies exhibited anchorage-independent growth, forming numerous colonies in soft agar assays. Molecular profiling demonstrated expression of several metastatic genes, but not of cadherin 1 (encoding the adhesion protein), in all patients. Conclusion: The authors successfully isolated, enumerated, and characterized CTCs from liquid biopsies of metastatic cancer patients. This study has potential to facilitate the development of new diagnostic and therapeutic methods using liquid biopsies, for application in metastatic cancers.
Key words:Circulating tumor cells,liquids biopsies,molecular markers,soft agar assay,metastasis,metastatic genes, epithelial mesenchymal transition, tumorigenic
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Journal of Cancer Metastasis and Treatment旨在分享癌症转移与治疗方面的最新研究，为该领域的研究学者和临床医生提供一个国际性学术交流平台。自2015年4月创刊以来，期刊读者遍布全球167个国家和地区，已在线发表文章125篇。诚挚欢迎您投稿!如有任何问题，请联系我们：editorialoffice@jcmtjournal.com
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