College of Life Sciences, Shaanxi Normal University, Xi'an, China.
Cancer Biother Radiopharm. 2010 Feb;25(1):55-64. doi: 10.1089/cbr.2008.0604.
Our previous work demonstrated that the enhancement of cytotoxicity by protoporphyrin IX (Pp) was significantly higher than hematoporphyrin (Hp), when they were compared at the same ultrasound-exposure conditions, since the cytoskeleton plays a crucial role in numerous cell functions and the effect of sonodynamic therapy (SDT) on cytoskeleton has not been reported yet. So, it is very important to investigate whether SDT has some influence on the cytoskeleton, and it would also be interesting to compare Pp-/ and Hp-mediated SDT on the cytoskeleton.
Ehrlich ascites carcinoma (EAC) cells were exposed to ultrasound for up to 15 seconds (1.34 MHz, 1 w/cm(2)) in the presence of 20 microM of Pp or 20 microM of Hp. Cell viability was determined by the trypan blue exclusion test. Morphologic changes were observed under a scanning electron microscope. Cytoskeleton damages were shown by immunofluorescence staining of microtubules, actin microfilaments, and intermediate filaments of vimentin.
Our experiment showed that the extent of cytoskeleton damage in EAC cells increased as time was prolonged, and also which was more significant in the Pp-SDT group than that in the Hp-SDT group at the same time after exposure.
Results implied that the ultrastructural changes might account for cell destruction and the cytoskeletal components were involved in the cell-damaging process induced SDT treatment.
我们之前的工作表明,在相同的超声暴露条件下,原卟啉 IX(Pp)的细胞毒性增强作用明显高于血卟啉(Hp),因为细胞骨架在许多细胞功能中起着至关重要的作用,而声动力学疗法(SDT)对细胞骨架的影响尚未见报道。因此,研究 SDT 是否对细胞骨架有一些影响非常重要,比较 Pp-和 Hp 介导的 SDT 对细胞骨架的影响也将非常有趣。
用 1.34MHz、1w/cm² 的超声在 20μM 的 Pp 或 20μM 的 Hp 存在下对艾氏腹水癌细胞(EAC)进行超声处理,时间长达 15 秒。用台盼蓝排斥试验测定细胞活力。用扫描电子显微镜观察形态变化。用微管、肌动蛋白微丝和中间丝波形蛋白的免疫荧光染色显示细胞骨架损伤。
我们的实验表明,随着时间的延长,EAC 细胞的细胞骨架损伤程度增加,在相同的暴露后时间内,Pp-SDT 组比 Hp-SDT 组更为显著。
结果表明,超微结构的变化可能导致细胞破坏,细胞骨架成分参与了 SDT 治疗诱导的细胞损伤过程。