转移，而不是原发肿瘤的生长，导致了大约90%的乳腺癌患者死亡。微触手（McTNs）的形成是转移的一个重要机制。三阴性乳腺癌（TNBC）是最具侵袭性的亚型，其靶向治疗有限。本研究旨在分离可行的循环肿瘤细胞（CTCs）并对其在药物治疗下进行功能分析。从20例TNBC患者中分离出CTCs，并在培养基中维持5天。免疫荧光染色和VyCap分析确定了生物标志物的表达。基于M30阳性细胞的检测评估了长春碱诱导的细胞凋亡。我们的研究结果显示，与患者细胞涂片中的CTC数目相比，使用TetherChips分析CTC绝对数目显著增加（p = 0.006）从而提供了足够的肿瘤细胞进行药物评估。长春碱处理（1小时）的活体CTC显著诱导细胞凋亡（p = 0.010）。它还导致Detyrosinated α-tubulin（GLU）和程序性死亡配体（PD-L1）表达的CTC显著减少（p < 0.001），并破坏了微触手。总之，这项初步研究提供了一种使用TetherChip技术进行活体CTC功能分析和药效评估的有用方案，为以患者个体水平针对转移性播散的靶向提供重要信息。

Metastasis, rather than the growth of the primary tumor, accounts for approximately 90% of breast cancer patient deaths. Microtentacles (McTNs) formation represents an important mechanism of metastasis. Triple-negative breast cancer (TNBC) is the most aggressive subtype with limited targeted therapies. The present study aimed to isolate viable circulating tumor cells (CTCs) and functionally analyze them in response to drug treatment. CTCs from 20 TNBC patients were isolated and maintained in culture for 5 days. Biomarker expression was identified by immunofluorescence staining and VyCap analysis. Vinorelbine-induced apoptosis was evaluated based on the detection of M30-positive cells. Our findings revealed that the CTC absolute number significantly increased using TetherChips analysis compared to the number of CTCs in patients' cytospins (p = 0.006) providing enough tumor cells for drug evaluation. Vinorelbine treatment (1 h) on live CTCs led to a significant induction of apoptosis (p = 0.010). It also caused a significant reduction in Detyrosinated α-tubulin (GLU), programmed death ligand (PD-L1)-expressing CTCs (p < 0.001), and disruption of McTNs. In conclusion, this pilot study offers a useful protocol using TetherChip technology for functional analysis and evaluation of drug efficacy in live CTCs, providing important information for targeting metastatic dissemination at a patient-individualized level.