Morphological Study on Permeating Efficiency and Localization of FCLA and HpD through Membrane of Lung Cancer Cell

摘要

It is reported that apoptosis of cancer cells in photodynamic therapy (PDT) is caused by ~1O_2 generated in photosensitization. In order to study the mechanism of this kind of ~1O_2-induced apoptosis, it is necessary to establish a special technique to dynamically detect intracellular production and localization of ~1O_2. FCLA, as a chemiluminescence probe to detect singlet oxygen(~1O_2) and superoxide (O_2~-), has been used successfully in photodynamic and sonodynamic diagnosis in tissue level, recently. This paper reported a preliminary result of morphological study on permeating efficiency and localization of FCLA and hematoporphyrin derivative (HpD) through cellular membrane. Human lung cancer cell line (ASTC-a-1) was used in the experiment. The result of this research showed that both HpD and FCLA could permeate through cellular membrane and localize to prinuclear area, when HpD or FCLA was incubated with cells. Although the molecular weight of HpD is close to FCLA's, the permeating efficiency of HpD through membrane was different from that of FCLA. Intracellular FCLA concentration reached a peak after incubation for only 30-45 minutes, but amount of HpD in cells approached the equilibrium after incubation for near 22 h. In the experiment, we did not observe the evidence of FCLA or HpD penetrating into nucleolus. This study suggests that it is possibly to use a specific chemiluminescence probe to dynamically detect the production and localization of ~1O_2 or O_2~- in cell.