基于BRET原理的肿瘤成像与可控治疗体系的构建及活性研究  

Construction and Activity Research of Tumor Imaging and Controllable Treatment System Based on BRET Principle

作　　者：谭湘玉 侯敏 李明达 罗妍洁 左悦 TAN Xiangyu;HOU Min;LI Mingda;LUO Yanjie;ZUO Yue(College of Physics and Chemistry,Hunan First Normal University,Changsha 410205,China)

机构地区：[1]湖南第一师范学院物理与化学学院,湖南长沙410205

出　　处：《生物化工》2024年第5期127-131,共5页Biological Chemical Engineering

基　　金：2022年度湖南省教育厅优秀青年项目(22B0874);2023年度湖南省大学生创新创业训练计划一般项目(S202312034040)。

摘　　要：本文旨在构建一种蛋白质-蛋白质相互作用体系,实现活体肿瘤生物成像和可控治疗肿瘤的目的。通过利用荧光素酶的催化生物发光特性,并结合孔雀石绿(MG)的荧光激活蛋白(FAP)可被光激活产生单线态氧的原理,构建了一种NanoLuc-FAP质粒。研究结果表明,NanoLuc的最大发射波长为460 nm,正好能够激发MG-FAP缀合物产生荧光,发射出的荧光最大发射波长为695 nm。此外,NanoLuc催化产生的荧光部分被FAP吸收并激活,其程度取决于MG的种类。激活后的FAP蛋白产生的单线态氧的量随着时间的增加而增加。MG的存在能够使NanoLuc的光信号增强。该研究设计出的基于生物发光共振能量转移的荧光素酶-荧光激活蛋白体系对于肿瘤成像和可控治疗起到一定的效果,对于今后进行临床监测和治疗具有参考意义。The aim of this paper is to construct a protein-protein interaction system for in vivo tumor bioimaging and controlled tumor treatment.A NanoLuc-FAP plasmid is constructed by exploiting the catalytic bioluminescent property of luciferase and combining with the fact that the fluorogen-activating protein(FAP)of malachite green(MG)can be lightactivated to produce single-linear oxygen.The results show that the maximum emission wavelength of NanoLuc is 460 nm,which is just enough to excite MG-FAP to produce fluorescence,and the maximum emission wavelength of the emitted fluorescence is 695 nm.In addition,the fluorescence produced by NanoLuc catalysis is partially absorbed and activated by FAP,and the degree of which depended on the species of MG.The amount of single-linear state oxygen produced by the activated FAP protein increase with time.The presence of MG is able to enhance the photoacoustic signal of NanoLuc.The bioluminescence resonance energy transfer-based luciferase-fluorescence-activated protein system designed in this study is effective for tumour imaging and controllable therapy,and will be useful for future clinical monitoring and therapy.

关 键 词：生物发光 荧光素酶 生物发光共振能量转移 荧光激活蛋白 单线态氧 

分 类 号：Q599[生物学—生物化学]