Carmona-Ule Nuria, González-Conde Miriam, Abuín Carmen, Cueva Juan F, Palacios Patricia, López-López Rafael, Costa Clotilde, Dávila-Ibáñez Ana Belén
Roche-Chus Joint Unit, Translational Medical Oncology Group, Oncomet, Health Research Institute of Santiago de Compostela (IDIS), Travesía da Choupana s/n, 15706 Santiago de Compostela, Spain.
CIBERONC, Centro de Investigación Biomédica en Red Cáncer, 28029 Madrid, Spain.
Cancers (Basel). 2021 May 28;13(11):2668. doi: 10.3390/cancers13112668.
Circulating tumor cells (CTC) have relevance as prognostic markers in breast cancer. However, the functional properties of CTCs or their molecular characterization have not been well-studied. Experimental models indicate that only a few cells can survive in the circulation and eventually metastasize. Thus, it is essential to identify these surviving cells capable of forming such metastases.
We isolated viable CTCs from 50 peripheral blood samples obtained from 35 patients with advanced metastatic breast cancer using RosetteSep for ex vivo culture. The CTCs were seeded and monitored on plates under low adherence conditions and with media supplemented with growth factors and Nanoemulsions. Phenotypic analysis was performed by immunofluorescence and gene expression analysis using RT-PCR and CTCs counting by the Cellsearch system.
We found that in 75% of samples the CTC cultures lasted more than 23 days, predicting a shorter Progression-Free Survival in these patients, independently of having ≥5 CTC by Cellsearch. We also observed that CTCs before and after culture showed a different gene expression profile.
the cultivability of CTCs is a predictive factor. Furthermore, the subset of cells capable of growing ex vivo show stem or mesenchymal features and may represent the CTC population with metastatic potential in vivo.
循环肿瘤细胞（CTC）作为乳腺癌的预后标志物具有重要意义。然而，CTC的功能特性或其分子特征尚未得到充分研究。实验模型表明，只有少数细胞能够在循环中存活并最终发生转移。因此，识别这些能够形成此类转移的存活细胞至关重要。
我们使用RosetteSep从35例晚期转移性乳腺癌患者的50份外周血样本中分离出活的CTC，用于体外培养。将CTC接种在低贴壁条件下的培养板上，并添加生长因子和纳米乳液的培养基进行监测。通过免疫荧光进行表型分析，使用RT-PCR进行基因表达分析，并通过Cellsearch系统对CTC进行计数。
我们发现，在75%的样本中，CTC培养持续超过23天，这预示着这些患者的无进展生存期较短，与通过Cellsearch检测到的≥5个CTC无关。我们还观察到培养前后的CTC显示出不同的基因表达谱。
CTC的可培养性是一个预测因素。此外，能够在体外生长的细胞亚群表现出干细胞或间充质特征，可能代表体内具有转移潜力的CTC群体。
