光敏剂HpD和单态氧探针FCLA的跨膜效率及其亚细胞定位研究

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

化学发光探针分子FCLA是一种海萤荧光素类似物分子,它可以选择性地与1O2及O2.反应产生化学发光,近年来已被成功用于在组织水平上进行光动力学和声动力学的肿瘤诊断中。但是FCLA在生物样品中能否进入细胞以及在细胞内的定位等问题目前尚不清楚。本文中报道利用激光共焦扫描显微镜进行FCLA和HpD的跨膜效率以及细胞内定位的形态学研究初步结果。结果表明,在37℃培养箱中用完全培养液进行培养时发现,HpD和FCLA都可以有效地跨膜,并定位在细胞质中。虽然FCLA与HpD的分子量大小相近,但是其进入肿瘤细胞的效率却并不相同。与HpD相比FCLA更容易进入细胞,对细胞没有明显的毒性。实验中未观测到FCLA和HpD进入细胞核的证据。本研究为利用1O2和O2.探针FCLA动态观测细胞内1O2或O2.的产生和定位建立了实验基础,并将推动在细胞或亚细胞水平上进行光动力学机制以及光敏过程引起细胞凋亡机制的研究。

FCLA, as a chemiluminescence probe to detect singlet oxygen（1^O2） and superoxide （O2^·）, has been used successfully in photodynamic and sonodynamic diagnosis in tissue level, recently. This paper reported a preliminary result of morphological study on permeating eftciency 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 one hour, but amount of HpD in cells approached an equilibrium after incubation for near 22 h. No significant ASIE-a-1 cytotoxity was detected at the FCLA doses of 0.1-100 μAM. 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 1^O2 or O2^·in cell.