Photodynamic effects of 5-aminolevulinic acid and its hexylester on several cell lines.

Shu-Min Wu, Qing-Guang Ren, Mi-Ou Zhou, Yi Wei, Ji-Yao Chen
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Abstract

5-aminolevulinic acid (ALA) and its hexyl-ester (He-ALA) has shown promising results in photodynamic detection and therapy of tumors. In this work, the photodynamic effects of ALA and He-ALA on neuroblastoma cells, hepatoma cells and fibroblast cells were comparatively studied. With the detection of fluorescence emission spectra, protoporphyrin IX (PpIX) induced by ALA or He-ALA was observed in these three cell lines. Confocal laser scanning microscope showed the diffuse PpIX fluorescence in cytoplasm of neuroblastoma cells. The kinetics of PpIX accumulation were different in these three kinds of cells. The PpIX content in hepatoma cells and fibroblast cells continuously increased with the incubation time of drugs until 12 h, while in neuroblastoma cells the PpIX content saturated around 8 h after incubation with ALA or He-ALA. In addition, the PpIX concentration in neuroblastoma cells was obviously higher than that in hepatoma cells and fibroblast cells, indicating that the PpIX production is cell line dependent. When incubated with ALA and irradiated with light, near 90% neuroblastoma cells were destroyed, while for hepatoma cells and fibroblast cells the death rate was around 50%. The results demonstrate that neuroblastoma cells are more sensitive to ALA-PDT and the neuro-tumor cells may be well suited for the treatment of ALA mediated photosensitization. Comparing to ALA, He-ALA can reach the similar results concerned PpIX production and PDT damaging in all three kinds of cells but with 10 times lower incubation concentration, demonstrating that He-ALA has higher efficiency than ALA on inactivation of cancer cells in vitro.

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5-氨基乙酰丙酸及其己酯对几种细胞系的光动力效应。
5-氨基乙酰丙酸(ALA)及其己基酯(He-ALA)在肿瘤的光动力学检测和治疗中显示出良好的效果。本研究比较研究了ALA和He-ALA对神经母细胞瘤细胞、肝癌细胞和成纤维细胞的光动力学作用。通过荧光发射光谱检测,在这三种细胞系中均观察到ALA或He-ALA诱导的原卟啉IX (PpIX)。共聚焦激光扫描显微镜下可见成神经细胞瘤细胞质中弥漫的PpIX荧光。PpIX的积累动力学在这三种细胞中不同。肝癌细胞和成纤维细胞中PpIX含量随着药物孵育时间的延长而不断增加,直至12 h,而神经母细胞瘤细胞中PpIX含量在ALA或He-ALA孵育后8 h左右达到饱和。此外,神经母细胞瘤细胞中PpIX的浓度明显高于肝癌细胞和成纤维细胞,表明PpIX的产生具有细胞系依赖性。用ALA孵育并光照时,近90%的神经母细胞瘤细胞被破坏,而肝癌细胞和成纤维细胞的死亡率约为50%。结果表明,神经母细胞瘤细胞对ALA- pdt更敏感,神经肿瘤细胞可能非常适合ALA介导的光敏治疗。与ALA相比,He-ALA在三种细胞中产生PpIX和破坏PDT的效果相似,但孵育浓度低10倍,表明He-ALA在体外对癌细胞的失活效率高于ALA。
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