Herrera-Ochoa Diego, Bravo Iván, Garzón-Ruiz Andrés
Departamento de Química Física, Facultad de Farmacia, Universidad de Castilla-La Mancha, Av. Dr. José María Sánchez Ibáñez, s/n, Albacete 02071, Spain.
Departamento de Química Física, Facultad de Farmacia, Universidad de Castilla-La Mancha, Av. Dr. José María Sánchez Ibáñez, s/n, Albacete 02071, Spain; Centro Regional de Investigaciones Biomédicas (CRIB), Unidad Asociada de Biomedicina (UCLM-CSIC), C/ Almansa, 14, Albacete 02008, Spain.
Colloids Surf B Biointerfaces. 2025 Jan;245:114265. doi: 10.1016/j.colsurfb.2024.114265. Epub 2024 Sep 21.
Anticancer therapies with cisplatin and volasertib (BI-6727) were monitored by fluorescence lifetime imaging microscopy (FLIM) in live SK-Mel-2 melanoma cells. A CdSe/ZnS quantum dot functionalized with a peptide containing D-penicillamine and histidine (CdSe/ZnS-PH) was used as intracellular pH fluorescent probe. A faster cytosol acidification was observed for cells treated with cisplatin when compared to volasertib. The first changes in the intracellular pH were found after 2 hours of treatment with cisplatin and 8 hours with volasertib. Additionally, the relationship between cytosol acidification and apoptosis was investigated using an innovative methodology based on time-resolved fluorescence measurements. Similar low percentages of apoptotic cells were observed after short incubation periods (2 - 8 hours) with both drugs. In contrast, late apoptosis and death were found for a large fraction of cells during 24-hour incubation with cisplatin but not volasertib. Thus, the early acidification observed in cisplatin treatment could accelerate apoptosis and cell death. Despite volasertib treatment shows slower mechanism of action than cisplatin, similar inhibitory effects were found for both drugs at longer incubation periods (72 hours). This study proves the potential of CdSe/ZnS-PH nanoparticle as a fluorescence lifetime probe in the study of the mechanism of action of antitumor drugs.
在活的SK-Mel-2黑色素瘤细胞中,通过荧光寿命成像显微镜(FLIM)监测了顺铂和沃拉替尼(BI-6727)的抗癌治疗效果。一种用含有D-青霉胺和组氨酸的肽功能化的CdSe/ZnS量子点(CdSe/ZnS-PH)被用作细胞内pH荧光探针。与沃拉替尼相比,用顺铂处理的细胞观察到更快的胞质酸化。在用顺铂处理2小时和用沃拉替尼处理8小时后发现细胞内pH的首次变化。此外,使用基于时间分辨荧光测量的创新方法研究了胞质酸化与细胞凋亡之间的关系。在用两种药物短时间孵育(2 - 8小时)后观察到相似的低凋亡细胞百分比。相比之下,在用顺铂而非沃拉替尼进行24小时孵育期间,发现大部分细胞发生晚期凋亡和死亡。因此,在顺铂治疗中观察到的早期酸化可能加速细胞凋亡和死亡。尽管沃拉替尼治疗显示出比顺铂更慢的作用机制,但在更长的孵育期(72小时)对两种药物发现了相似的抑制作用。这项研究证明了CdSe/ZnS-PH纳米颗粒作为荧光寿命探针在抗肿瘤药物作用机制研究中的潜力。