Zhang Lei, Lei Jianping, Liu Jintong, Ma Fengjiao, Ju Huangxian
State Key Laboratory of Analytical Chemistry for Life Science , School of Chemistry and Chemical Engineering , Nanjing University , Nanjing 210093 , P. R. China . Email:
Chem Sci. 2015 Jun 1;6(6):3365-3372. doi: 10.1039/c5sc00471c. Epub 2015 Mar 26.
The evolution of the intracellular caspase family is crucial in cell apoptosis. To evaluate this process, a universal platform of activation and monitoring of the evolution of intracellular caspase is designed. Using well-known gold nanostructure as a model of both nanocarrier and matter inducing the cell apoptosis for photothermal therapy, a nanoprobe is prepared by assembly of two kinds of dye-labelled peptides specific to upstream caspase-9 and downstream caspase-3 as the signal switch, and folic acid as a targeting moiety. The energy transfer from dyes to the gold nanocarrier at two surface plasmon resonance absorption wavelengths leads to their fluorescence quenching. Upon endocytosis of the nanoprobe to perform the therapy against cancer cells, the peptides are successively cleaved by intracellular caspase activation with the evolution from upstream to downstream, which lights up the fluorescence of the dyes sequentially, and can be used to quantify both caspase-9 and caspase-3 activities in cancer cells and to monitor their evolution in living mice. The recovered fluorescence could also be used to assess therapeutic efficiency. This work provides a novel powerful tool for studying the evolution of the intracellular caspase family and elucidating the biological roles of caspases in cancer cell apoptosis.
细胞内半胱天冬酶家族的演变在细胞凋亡过程中至关重要。为评估这一过程,设计了一个用于激活和监测细胞内半胱天冬酶演变的通用平台。以著名的金纳米结构作为纳米载体和诱导光热疗法细胞凋亡的物质模型,通过组装两种分别针对上游半胱天冬酶 -9 和下游半胱天冬酶 -3 的染料标记肽作为信号开关,以及叶酸作为靶向部分,制备了一种纳米探针。在两个表面等离子体共振吸收波长处,染料向金纳米载体的能量转移导致其荧光猝灭。纳米探针通过胞吞作用对癌细胞进行治疗时,肽会随着从上游到下游的演变被细胞内半胱天冬酶激活依次切割,从而使染料的荧光依次亮起,可用于定量癌细胞中半胱天冬酶 -9 和半胱天冬酶 -3 的活性,并监测它们在活体小鼠中的演变。恢复的荧光还可用于评估治疗效果。这项工作为研究细胞内半胱天冬酶家族的演变以及阐明半胱天冬酶在癌细胞凋亡中的生物学作用提供了一种新型强大工具。