具有聚集诱导发光特性的线粒体靶向荧光探针在骨肿瘤诊疗中的应用

Application of mitochondria targeting probes with aggregation-induced emission properties in the diagnosis and treatment of bone tumors

目的探讨MeTTPy的聚集诱导发光(AIE)性质,细胞器靶向性,单光子成像效果,以及光动力治疗(PDT)效果。方法采用人源骨肉瘤细胞系MG63细胞作为细胞模型,探讨MeTTPy浓度为1μmol/L时的生物相容性,以商业线粒体染料为比较对象,探讨其亚细胞水平的靶向区域和靶向能力,并探讨其活性氧(ROS)产生能力,应用28 mw/cm^(2)白光光源辐照,探讨其不同浓度对肿瘤细胞的杀伤率,应用活死细胞染色法评价MeTTPy对MG63肿瘤细胞的潜在的光动力治疗效果。采用t检验确定治疗组与对照组之间的显著性。结果MeTTPy浓度为1μmol/L时具有良好的生物相容性,其对线粒体的靶向性能与商业线粒体染料具有90%的共染率,对活细胞中的线粒体具有高特异性靶向;MeTTPy具有较强的原位ROS产生能力,其ROS产生量与对照组比较明显增加;细胞毒性测试实验显示,与对照组MG63细胞存活率比较,MeTTPy浓度为2.5μmol/L时MG63细胞存活率不受影响(t=1.862,P>0.05),差异无统计学意义,表现出其良好的生物相容性。采用低功率白光光源照射下(28 mw/cm^(2)),MeTTPy在较低的浓度下(2.5μmol/L),MG63细胞可被大比例杀伤(杀伤率>92%,t=16.35,P<0.05),显示出MeTTPy对肿瘤细胞具有良好的光疗效果。结论MeTTPy具有良好的荧光成像功能,对线粒体具有高靶向性,具有高效率的ROS产生能力,能够对骨肉瘤细胞进行高效杀伤,是一款高效的光敏剂,可以作为骨肉瘤等肿瘤细胞进行手术切除等治疗方式的良好辅助。

Objective To investigate the aggregation-induced luminescence(AIE)properties,organelle targeting,single photon imaging and photodynamic therapy(PDT)effects of MeTTPy.Methods Human osteosarcoma cell line MG63 was used as a cell model to explore the biocompatibility of MeTTPy at a concentration of 1μmol/L.Commercial mitochondrial dyes were used as a comparison object to explore the targeting region and targeting ability of MeTTPy at the subcellular level,and to discuss reactive oxygen species.The killing rate of different concentrations of MeTTPy on tumor cells was investigated by 28 mw/cm^(2) white light source irradiation,t test was used to determine the significance between the treatment group and the control group,and the potential photodynamic therapeutic effect of MeTTPy on MG63 tumor cells was evaluated by live and dead cell staining.Results MeTTPy had good biocompatibility,and its targeting performance to mitochondria had 90%co-staining rate with commercial mitochondrial dyes.MeTTPy was highly specific to mitochondria in living cells.It had strong ability to produce reactive oxygen species(ROS)in situ.Cytotoxicity test showed that compared with the control group,MG63 cell survival rate was not affected when MeTTPy concentration was 2.5μmol/L(t=1.862,P>0.05),and the difference was not statistically significant,indicating good biocompatibility.Under the irradiation of low power white light source(28 mw/cm^(2)),MG63 cells could be killed in a large proportion(killing rate>92%)at a low concentration(2.5μmol/L),indicating that MeTTPy had a good phototherapy effect on tumor cells.Conclusion MeTTPy has good aggregation-induced luminescence performance,has high targeting to mitochondria without the need of carriers such as wrapping,and has a high efficiency ROS generation ability,which can provide efficient fluorescence image-guided photodynamic therapy effect.