摘要
目的:探讨血卟啉单甲醚(HMME)介导的光动力疗法(HMME-PDT)对HL60细胞的作用及PDT前后HL60细胞表面超微结构的变化。方法:CCK-8法检测光敏剂浓度和光照剂量对HL60细胞抑制率的影响,荧光分光光度计监测PDT过程中光敏剂荧光强度随时间的变化,Fluo 3-AM荧光探针检测不同浓度HMME作用后HL60细胞内Ca2+变化,原子力显微镜观测PDT作用前后不同扫描范围HL60细胞表面的超微结构图。结果:细胞灭活率呈光敏剂浓度-光剂量依赖关系,当HMME为50μg/mL,光照剂量为24 J/cm2时,灭活效率达到70%;随着光照时间的增加,光敏剂的荧光强度不断减弱,下降速率也逐渐变慢;随HMME作用浓度增加,钙离子浓度显著升高;HMME-PDT作用后HL60细胞表面结构出现明显变化。结论:HMME-PDT能有效灭活HL60细胞,光敏剂浓度和光剂量是影响PDT疗效的重要因素,PDT过程中伴随有光漂白现象的发生,细胞凋亡和钙离子浓度增加呈正相关,PDT作用前后细胞出现明显萎缩,细胞膜粗糙度增加。
Objective： To investigate the effectiveness of HMME-mediated PDT on HL60 cells and the morphological changes of HL60 due to the PDT process. Methods： CCK-8 was used to evaluate the influence of HMME concentration and light dosage on the inhibition ratio to HL60, while the changes of fluorescence intensity of HMME were detected by fluorescence spectrophotometer over time. Fluo 3-AM was used to measure the variation of Ca^2＋ after PDT with different concentration of HMME（ Cells were stained by Fluo 3-AM and the change of Ca^2＋ resulted from different concentration of HMME was determined）. The morphological changes of HL60 cells were examined by AFM（ Atomic Force Microsco- py）. Results：The inhibition rate exhibits HMME concentration and light dosage-dependent manner. Fluorescence intensity decreased and the decrease rate showed down over time, when the concentration of Ca^2＋ increased. Additionally the surface images obtained by AFM showed obvious morphological changes. Conclusions：HMME-PDT could effectively in- duce the death of HL60 cells meanwhile concentration of HMME and light dosage are key factors involved in PDT. Pho- tobleaching of HMME was observed in the PDT process. Apoptosis of HL60 was positively related to the concentration of Ca^2＋ . Cells became atrophy and the roughness of membrane increased after PDT.
出处
《激光生物学报》
CAS
CSCD
2012年第4期304-309,共6页
Acta Laser Biology Sinica
基金
国家自然科学基金项目(61072029)
广东省自然科学基金项目(10151063101000025)
广州市科技计划项目(2010Y1-C111)
