Monitoring the Cellular Delivery of Doxorubicin-Cu Complexes in Cells by Fluorescence Lifetime Imaging Microscopy
Monitoring the Cellular Delivery of Doxorubicin-Cu Complexes in Cells by Fluorescence Lifetime Imaging Microscopy
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通过荧光寿命成像显微镜监测细胞中阿霉素-铜复合物的细胞递送
DOI：
10.1021/acs.jpca.0c00182
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发表时间：
2020
影响因子：
2.9
通讯作者：
Qu Junle
中科院分区：
文献类型：
--
作者：
Peng Zheng;Nie Kaixuan;Song Yiwan;Liu Hao;Zhou Yingxin;Yuan Yufeng;Chen Danni;Peng Xiao;Yan Wei;Song Jun;Qu Junle
在药物前研究领域，在药物效应研究中，从传统的终点生化分析中获得的信息既不能提供单个细胞的动态过程，也不能提供单个细胞的异质性反应。成像显微镜技术，特别是荧光寿命成像显微镜（FLIM），可以满足这些要求。在这项工作中，我们使用FLIM技术观察了阿霉素(Dox) -Cu复合物在KYSE150活细胞中的进入和释放。Dox - cu配合物的荧光信号较弱，但寿命值与原始Dox相近，其荧光可以通过添加生物硫醇化合物（如谷胱甘肽）释放。细胞活力实验结果表明，Dox-Cu化合物对KYSE150细胞具有良好的杀伤作用。FLIM数据显示，KYSE150细胞细胞质中的Dox-Cu复合物释放出游离阿霉素，并在细胞核中积累。给药90 min后，细胞质和细胞核中的荧光寿命信号分别达到1.21和1.46 ns，反映了Dox-Cu复合物的转化和运输。本研究为FLIM技术监测前药的研究提供了良好的范例，拓展了FLIM技术在药物开发中的有益应用。
In the prodrug research field, information obtained from traditional end point biochemical assays in drug effect studies could provide neither the dynamic processes nor heterogeneous responses of individual cells.In situimaging microscopy techniques, especially fluorescence lifetime imaging microscopy (FLIM), could fulfill these requirements. In this work, we used FLIM techniques to observe the entry and release of doxorubicin (Dox)–Cu complexes in live KYSE150 cells. The Dox–Cu complex has weaker fluorescence signals but similar lifetime values as compared to the raw Dox, whose fluorescence could be released by the addition of biothiol compound (such as glutathione). The cell viability results indicated that the Dox–Cu compound has a satisfactory killing effect on KYSE150 cells. The FLIM data showed that free doxorubicin was released from Dox–Cu complexes in cytoplasm of KYSE150 cells and then accumulated in the nucleus. After 90 min administration, the fluorescence lifetime signals reached 1.21 and 1.46 ns in the cytoplasm and nucleus, respectively, reflecting the transformation and transportation of Dox–Cu complexes. In conclusion, this work provides a satisfactory example for the research of prodrug monitored by FLIM techniques, expanding the useful applications of FLIM technique in drug development.