Gala de Pablo Julia, Chisholm David R, Ambler Carrie A, Peyman Sally A, Whiting Andrew, Evans Stephen D
Molecular and Nanoscale Physics Group, School of Physics and Astronomy, University of Leeds, Leeds, UK.
Analyst. 2020 Aug 24;145(17):5878-5888. doi: 10.1039/d0an01023e.
Raman spectroscopy has been used to observe uptake, metabolism and response of single-cells to drugs. Photodynamic therapy is based on the use of light, a photosensitiser and oxygen to destroy tumour tissue. Here, we used single-cell Raman spectroscopy to study the uptake and intracellular degradation of a novel photosensitiser with a diphenylacetylene structure, DC473, in live single-cells from colorectal adenocarcinoma cell lines SW480, HT29 and SW620. DC473 was seen to predominantly accumulate in lipid droplets, showing higher accumulation in HT29 and SW620 cells than in SW480 cells, with a broader DC473 peak shifted to higher wavenumbers. DC473 activation and effects were tracked on live single-cells for 5 minutes. Upon exposure to UV light, the DC473 signal intensity dropped, with remaining DC473 shifting towards higher wavenumbers and widening, with a lifetime of approximately 50 seconds. Morphologically, SW480 and SW620 cells showed changes upon photodynamic therapy, whereas HT29 cells showed no changes. Morphological changes correlated with higher remaining DC473 signal after UV exposure. Our research suggests that DC473 forms aggregates within the cells that disaggregate following activation, showing the potential of Raman spectroscopy for the study of time-dependent single-cell pharmacodynamics.
拉曼光谱已被用于观察单细胞对药物的摄取、代谢及反应。光动力疗法基于利用光、光敏剂和氧气来破坏肿瘤组织。在此，我们使用单细胞拉曼光谱研究了一种具有二苯乙炔结构的新型光敏剂DC473在结肠直肠腺癌细胞系SW480、HT29和SW620的活单细胞中的摄取及细胞内降解情况。观察到DC473主要积聚在脂滴中，在HT29和SW620细胞中的积聚量高于SW480细胞，且DC473的峰更宽并向更高波数移动。在活单细胞上追踪DC473的激活及效应持续5分钟。暴露于紫外光后，DC473信号强度下降，剩余的DC473向更高波数移动并变宽，其寿命约为50秒。形态学上，光动力疗法后SW480和SW620细胞出现变化，而HT29细胞未出现变化。形态学变化与紫外光照射后更高的剩余DC473信号相关。我们的研究表明，DC473在细胞内形成聚集体，激活后解聚，这显示了拉曼光谱在研究时间依赖性单细胞药效学方面的潜力。
