Fernandez-Carrascal Ana, Garcia-Algar Manuel, Nazarenus Moritz, Torres-Nuñez Alicia, Guerrini Luca, Feliu Neus, Parak Wolfgang J, Garcia-Rico Eduardo, Alvarez-Puebla Ramon A
Department of Physical Chemistry and EMaS, Universitat Rovira i Virgili, Tarragona, Spain.
Karolinska Institutet, Stockholm, Sweden.
Oncotarget. 2017 Jun 16;8(44):76108-76115. doi: 10.18632/oncotarget.18551. eCollection 2017 Sep 29.
Quantification of circulating tumor cells (CTCs) in blood samples from cancer patients is a non-invasive approach to monitoring the status of the disease. Most of the methods proposed in the recent years are phenomenological and rely on the use of antibodies labelled with fluorophores, magnetic particles, or immobilized on surfaces to capture the CTCs. Herein, we designed and optimized a method that employs a glucose analogue labelled with a fluorophore which takes advantage of the different metabolic pathways of cancer cells to discern them from normal ones. Notably, we demonstrate that fluorescence signal in tumor cells can be greatly maximized by applying hyperoxia conditions without damaging the cells. These results are demonstrated by means of confocal fluorescence and flow-cytometry measurements in peripheral blood mononuclear cells (PBMC) extracted after Ficoll of human blood samples and spiked with a known concentration of MCF-7 tumor cells.
对癌症患者血液样本中的循环肿瘤细胞(CTC)进行定量分析是一种监测疾病状态的非侵入性方法。近年来提出的大多数方法都是现象学的,依赖于使用标记有荧光团、磁性颗粒或固定在表面的抗体来捕获CTC。在此,我们设计并优化了一种方法,该方法采用标记有荧光团的葡萄糖类似物,利用癌细胞不同的代谢途径将其与正常细胞区分开来。值得注意的是,我们证明了在不损害细胞的情况下,通过应用高氧条件可以极大地增强肿瘤细胞中的荧光信号。这些结果通过对人血样本进行Ficoll分离后提取的外周血单核细胞(PBMC)进行共聚焦荧光和流式细胞术测量得到了证实,这些PBMC中加入了已知浓度的MCF-7肿瘤细胞。