超声结合原卟啉Ⅸ对腹水型S180肿瘤细胞的损伤作用

S180 cell damage induced by ultrasonically-activated protoporphyrin Ⅸ disodium salt

目的:根据原卟啉IX(PpIX)在S180肿瘤细胞内的含量变化及亚细胞定位分析,研究聚焦超声结合PpIX对S180肿瘤细胞的损伤作用。方法:应用荧光分光光度法测定PpIX在S180肿瘤细胞内的富集;激光共聚焦扫描显微镜观察PpIX的亚细胞定位;台盼蓝拒染法检测超声结合不同浓度的PpIX作用后细胞的存活率;环境扫描电镜观察细胞膜表面超微结构变化。结果:PpIX在S180肿瘤细胞内的富集是一个动态变化过程,45min达到最高点,此时为最佳声照处理时间点,且此时PpIX主要定位于细胞膜;超声激活PpIX对S180肿瘤细胞的杀伤作用明显大于单纯超声组,其膜损伤程度也更为严重。结论:细胞膜可能是超声激活PpIX作用的一个主要靶点。

Objective： To study killing effect on sarcoma 180 by the combination of PpⅨ and ultrasound, according to the accurate subcellular localization of PpⅨ and the change of its concentration in S180 cell after administration. Methods： The cellular accumulation of PpⅨ in S180 was estimated by fluorence spectrophotometer. Laser scanning confocal microscope observation was performed to study the subcellular location pattern of PpⅨ. Cell viability was evaluated by trypan blue exclution and morphological changes were evaluated by means of scanning electron microscope. Results： The accumulation of PpⅨ in S180 was a kinetic process, and achieved its most content at 45 min, which was the better exposure time, when PpⅨ was mainly distributed in cell membrane. Ultrasound in the presence of PpⅨ induced a more antitumor effect than only ultrasound did, and the damage of membrane was more serious. Conclusion： The cell membrane may be one of the most important targets for ultrasonically-activated PpⅨ.