Laboratory of Biochemistry/Metastatic Signaling, Section of Genetics, Cell Biology and Development, Department of Biology, University of Patras, GR-26504 Patras, Greece.
Department of Pharmacology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Cells. 2023 Jul 26;12(15):1940. doi: 10.3390/cells12151940.
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 ( = 0.006) providing enough tumor cells for drug evaluation. Vinorelbine treatment (1 h) on live CTCs led to a significant induction of apoptosis ( = 0.010). It also caused a significant reduction in Detyrosinated α-tubulin (GLU), programmed death ligand (PD-L1)-expressing CTCs ( < 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.
转移而非原发性肿瘤的生长,约占 90%乳腺癌患者的死亡原因。微刺突(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,靶向治疗有限。本研究旨在分离有活力的循环肿瘤细胞(CTC)并对其进行功能分析以响应药物治疗。从 20 名 TNBC 患者中分离并在培养中维持 CTC 5 天。通过免疫荧光染色和 VyCap 分析鉴定生物标志物表达。基于 M30-阳性细胞的检测评估长春瑞滨诱导的细胞凋亡。我们的研究结果表明,与患者细胞涂片上的 CTC 数量相比( = 0.006),TetherChips 分析显著增加了 CTC 的绝对数量,为药物评估提供了足够的肿瘤细胞。长春瑞滨(1 h)处理活 CTC 导致细胞凋亡显著诱导( = 0.010)。它还导致去酪氨酸化的 α-微管蛋白(GLU)、程序性死亡配体(PD-L1)表达的 CTC 显著减少( < 0.001),并且微刺突破坏。总之,这项初步研究提供了一种使用 TetherChip 技术对活 CTC 进行功能分析和药物疗效评估的有用方案,为在个体化患者水平上靶向转移扩散提供了重要信息。