摘要
目的:通过检测超声加热处理后Tca8113细胞凋亡的主要相关参数变化,探讨超声加热诱导肿瘤细胞凋亡的机制。方法:应用超声热辐射系统,分别对Tca8113细胞进行不同温度、时间的加热处理,流式细胞仪连续检测42℃超声加热10min后Tca8113细胞早期凋亡和继发性坏死的动态变化,以及不同处理组的线粒体跨膜电位(ΔΨm)、Caspase-3活性的变化。采用SAS6.12统计软件包进行方差分析,比较各组均数间的显著性水平。结果:42℃超声加热(10min)Tca8113细胞后1h,即检测到细胞早期凋亡,6~8h达到高峰,以后迅速下降,12h后保持在较低水平,细胞的继发性坏死伴随着凋亡出现,在高水平保持一段时间,10h后开始下降。其与早期凋亡呈正相关(r=0.7909,P=0.0064);无论温度梯度(38℃~44℃,10min)还是时间梯度(42℃,10min~60min)的超声加热,均能导致ΔΨm显著降低(P<0.01)、Caspase-3活性显著升高(P<0.01),且两者具有显著相关性(温度梯度组r=0.89189,P=0.0029,时间梯度组r=0.9679,P=0.0003)。结论:超声加热能诱导Tca8113细胞凋亡,导致ΔΨm降低、Caspase-3活性水平升高。说明超声加热通过线粒体-Caspase途径,诱导Tca8113细胞凋亡。
PURPOSE: To investigate the mechanism of apoptosis in Tca8113 cells induced by ultrasound hyperthermia by detecting changes in related index. METHODS: Tca8113 cells were treated in vitro by ultrasound hyperthermia in different heating temperatures (38℃ to 44℃,10 minutes) and heating times (42℃,10 to 60 minutes), and then the dynamic changes of early apoptosis and secondary necrosis treated by 42℃ for 10 minutes were assayed to detect A^m and Caspase-3 levels of different groups by flow cytometry (FCM). The means of each group were compared by ANOVA with SAS6.12 software package. RESULTS: After heated by ultrasound in 42℃ for 10 minutes, the early apoptosis of Tca8113 was detected. The apoptosis index reached its highest level at the 6^th to 8^th hour, then decreased rapidly and maintained in a lower level after 12 hours.The level of secondary necrosis increased with the level of early apoptosis, but kept in a higher level until the 10^th hour, the level of secondary necrosis correlated with that of the early apoptosis(r=0.7909, P=0.0064). The fraction of cells with low mitochondria membrane potential and increased activity of Caspase-3 were detected either in the heating-temperature grads group or in the heating-time grads group, which showed significant relationship between the two apoptosis related indes (r=0.89189,P=0.0029 in thd heating-tempcrature grads groups,r=0.9679,P=0.0003 in the heating-time grads group).CONCLUSION:Tea8113 cells devcloped apoptosis after heated by ultmsound byperthermin.along with △ψm leyel decneasing and Caspase-3 aetivity inereasing.Ultrasound byperthermia induees apoplosis of Tca8113 cells by thd mitochomlmm-Caspase pathway.
出处
《上海口腔医学》
CAS
CSCD
2006年第5期507-511,共5页
Shanghai Journal of Stomatology
基金
国家自然科学基金(30471898)
上海市科学技术委员会科研基金(041111041)
上海交通大学-上海第二医科大学合作基金(03-02)~~