血卟啉甲醚声动力诱导体外C6胶质瘤细胞凋亡与蛋白Bcl-2/Bax表达

Apoptosis by sonodynamic therapy with hematoporphyrin monomethyl ether in C6 glioma cells in vitro and protein expression of Bcl-2/Bax

目的探讨血卟啉甲醚声动力疗法(sonodynamic theraoy,SDT)对体外C6胶质瘤细胞的促凋亡作用及蛋白Bcl-2/Bax表达。方法取处于对数生长期C6胶质瘤细胞接种于96孔6孔培养板上。采用噻唑蓝比色分析法(MTT)观察不同时间超声辐照后细胞存活率,筛选最佳照射时间;流式细胞仪检测24h后对照组(control)、血卟啉甲醚组(hematoporphyrin monomethyl ether,HMME)、单纯超声组(ultrasound)、超声联合HMME组(SDT)C6胶质瘤细胞凋亡率;Western blot检测凋亡蛋白Bcl-2、Bax的表达。结果当超声频率为1.0MHz,声强为1.0W/cm2,MTT筛选60s为最佳辐照时间。同对照组、HMME及单纯超声组比较,SDT组凋亡率明显增高(P<0.05),同时伴有凋亡蛋白Bax,上调表达,Bcl-2下调表达。结论血卟啉甲醚声动力诱导体外C6胶质瘤细胞凋亡,Bcl-2/Bax参与并调节凋亡过程。

Objective To study apoptotic effect of SDT with hematoporphyrin monomethyl ether （HMME） on C6 glioma cells in vitro and relation with protein expression of Bcl-2/Bax.Methods Time of ultrasound irradiation was optimized by MTr. Apoptotic rates were detected by flow cytometry, and protein expression of Bcl-2, Bax were measured by Western blot after SDT treatment. Resuits Treatment of C6 glioma cells with SDT resulted in the occurrence of apoptosis. In the apoptosis, expression of Bax was significantly increased, and expression of Bcl-2 was also significantly decreased after SDT treatment. Conclusion Ultrasound with HMME could induce apoptosisi in C6 glioma cells in vitro, and the apoptosis meidiatied by SDT was associated with and regulated by protein Bcl-2/Bax .