利用微流控 Western 印迹技术分析循环肿瘤细胞中的蛋白质表达。

Profiling protein expression in circulating tumour cells using microfluidic western blotting.

机构信息

Department of Bioengineering, University of California, Berkeley, 308B Stanley Hall, MC 1762, Berkeley, California 94720, USA.

Vortex Biosciences, Inc., Menlo Park, California 94025, USA.

出版信息

Nat Commun. 2017 Mar 23;8:14622. doi: 10.1038/ncomms14622.

DOI:10.1038/ncomms14622

PMID:28332571

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5376644/

Abstract

Circulating tumour cells (CTCs) are rare tumour cells found in the circulatory system of certain cancer patients. The clinical and functional significance of CTCs is still under investigation. Protein profiling of CTCs would complement the recent advances in enumeration, transcriptomic and genomic characterization of these rare cells and help define their characteristics. Here we describe a microfluidic western blot for an eight-plex protein panel for individual CTCs derived from estrogen receptor-positive (ER+) breast cancer patients. The precision handling and analysis reveals a capacity to assay sparingly available patient-derived CTCs, a biophysical CTC phenotype more lysis-resistant than breast cancer cell lines, a capacity to report protein expression on a per CTC basis and two statistically distinct GAPDH subpopulations within the patient-derived CTCs. Targeted single-CTC proteomics with the capacity for archivable, multiplexed protein analysis offers a unique, complementary taxonomy for understanding CTC biology and ascertaining clinical impact.

摘要

循环肿瘤细胞（CTCs）是在某些癌症患者的循环系统中发现的稀有肿瘤细胞。CTCs 的临床和功能意义仍在研究中。CTCs 的蛋白质谱分析将补充最近在这些稀有细胞的计数、转录组和基因组特征描述方面的进展，并有助于定义它们的特征。在这里，我们描述了一种用于从雌激素受体阳性（ER+）乳腺癌患者中分离的单个 CTC 的八重蛋白面板的微流控 Western blot。精确的处理和分析显示了检测有限的患者来源 CTC 的能力，一种比乳腺癌细胞系更具抗裂解性的生物物理 CTC 表型，一种能够基于每个 CTC 报告蛋白表达的能力，以及在患者来源的 CTC 内两个统计学上不同的 GAPDH 亚群。具有存档、多重蛋白分析能力的靶向单个 CTC 蛋白质组学为理解 CTC 生物学和确定临床影响提供了一种独特的、互补的分类法。