超声激活血卟啉诱导艾氏腹水瘤细胞凋亡的细胞学研究

Study on apoptosis of enrlich ascites tumor cells induced by ultrasoundly activated hematoporphyrin

采用频率为1.34 MHz、声强为1W/cm2的超声结合血卟啉对EAT细胞凋亡模式进行了研究,探讨了声动力学疗法抗肿瘤的分子机制.超声激活血卟啉作用于EAT细胞后,不同时间点取材,通过扫描电镜观察EAT细胞的形态学变化;通过HO荧光染色观察凋亡细胞的形态学特征;利用免疫细胞化学方法检测caspase-8蛋白的活性变化.结果表明,超声激活血卟啉可诱导EAT细胞凋亡的发生,并且随时间的延迟凋亡现象愈加明显;免疫细胞化学方法表明超声处理后caspase-8蛋白表达活性显著增强,且于1h活化程度达到最高.研究表明,超声激活血卟啉可诱导EAT细胞凋亡,其作用的分子机制可能涉及Caspase-8依赖的膜受体凋亡信号调节通路.

The effect of ultrasound with a frequency of 1.34 MHz and intensity of 1W/cm^2 in presence of hematoporphyrin on EAT tumor cells was studied. The changes in the ultrastructure of the samples obtained at different time points were observed by the scanning electron microscope. HO apoptosis detecting kit was used to identify the morphologic changes of EAT cells after the ultrasound treatment at different time points. The expression of apoptosis associated protein Caspase-8 were detected by immunohistochemistry techniques. The data shown that the apoptosis in EAT tumor cells can be induced by ultrasonically activated hematoporphyrin, and the apoptosis was enhanced gradually by the ultrasounic treatment. The activity of Caspase-8 were apparently up-regulated during apoptosis. The results above indicate that sonochemically activated hematoporphyrin mediates the apoptosis of tumor cells,and the mechanism may be related to the expression of Caspase-8 cleaving function depended on membrane receptor activate on pathways.