Characterization of endogenous fluorescence in nonsmall lung cancerous cells: A comparison with nonmalignant lung normal cells
Characterization of endogenous fluorescence in nonsmall lung cancerous cells: A comparison with nonmalignant lung normal cells
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DOI：
10.1002/jbio.201960210
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发表时间：
2020-02-26
影响因子：
2.8
通讯作者：
Ohta, Nobuhiro
中科院分区：
文献类型：
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作者：
Awasthi, Kamlesh;Chang, Feng-Lin;Ohta, Nobuhiro
在正常和癌细胞中监测内源性荧光团如烟酰胺腺嘌呤二核苷酸（NADH）和黄素腺嘌呤二核苷酸（FAD）的荧光性质提供了关于癌细胞代谢功能障碍的生物化学和生物物理方面的非侵入性信息。本文报道了人肺非小细胞癌细胞（H661和A549）和正常肺细胞（MRC-5）中NADH和FAD的时间分辨光谱和荧光寿命图像。两种荧光团在脉冲激发时显示快速和缓慢衰减的发射分量，并且NADH和FAD的荧光光谱在其衰减期间分别显示蓝移和红移。发现癌细胞中所有已鉴定的NADH和FAD的寿命组分均短于正常细胞，无论在不同的细胞外条件下（细胞悬浮在比色皿中，细胞附着在玻璃基底上）如何测量，这表明代谢的变化可能改变了亚细胞环境，并可能影响NADH和FAD与这些辅因子结合的酶的相互作用。癌细胞的NADH和FAD的强度比也大于正常细胞。
Monitoring fluorescence properties of endogenous fluorophores such as nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) in normal and cancerous cells provide substantial information noninvasively on biochemical and biophysical aspects of metabolic dysfunction of cancerous cells. Time-resolved spectral profiles and fluorescence lifetime images of NADH and FAD were obtained in human lung nonsmall carcinomas (H661 and A549) and normal lung cells (MRC-5). Both fluorophores show the fast and slowly decaying emission components upon pulsed excitation, and fluorescence spectra of NADH and FAD show blue- and red-shifts, respectively, during their decay. All identified lifetime components of NADH and FAD were found to be shorter in cancerous cells than in normal cells, no matter how they were measured under different extra-cellular conditions (cells suspended in cuvette and cells attached on glass substrate), indicating that the changes in metabolism likely altered the subcellular milieu and potentially also affected the interaction of NADH and FAD with enzymes to which these cofactors were bound. The intensity ratio of NADH and FAD of cancerous cells was also shown to be larger than that of normal cells.