In vitro study of low intensity ultrasound combined with different doses of PDT: Effects on C6 glioma cells.
In vitro study of low intensity ultrasound combined with different doses of PDT: Effects on C6 glioma cells.
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DOI：
10.3892/ol.2012.1060
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发表时间：
2013-02
期刊：
影响因子：
2.9
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Li JH;Chen ZQ;Huang Z;Zhan Q;Ren FB;Liu JY;Yue W;Wang Z
本研究旨在探讨血卟啉单甲醚（HMME）介导的声动力疗法联合光动力疗法（SPDT）对C6胶质瘤细胞的杀伤作用。在SPDT组中，以0.5W/cm 2的强度和1 MHz的频率用超声处理细胞，随后进行不同剂量的光照射。MTT法测定细胞生长抑制率。流式细胞仪检测细胞凋亡率。通过免疫印迹研究胱天蛋白酶3、8和9的切割。通过荧光酶标仪测量活性氧（ROS）。SPDT对胶质瘤细胞的作用也在各种ROS清除剂存在或不存在的情况下进行了研究。在光剂量<200 J/cm 2时，SPDT对C6胶质瘤细胞的生长抑制率明显高于单纯声动力学疗法（SDT）或光动力学疗法（PDT）。当光剂量增加到>120 J/cm 2时，SPDT处理的C6胶质瘤细胞的生长抑制率没有显著升高。光剂量为80 J/cm 2时，SPDT组细胞凋亡率最高。SPDT组产生的ROS量明显高于SDT组和PDT组。添加NaN 3或甘露醇可降低SPDT的生长抑制率。总之，我们的数据表明，SPDT在体外通过SDT和PDT的协同作用，有力地杀死C6胶质瘤细胞。PDT诱导C6胶质瘤细胞凋亡的途径包括线粒体途径和死亡受体途径。此外，ROS可能在SPDT中起重要作用。
The aim of this study was to study the effects of killing C6 glioma cells induced by hematoporphyrin monomethyl ether (HMME)-mediated sonodynamic therapy combined with photodynamic therapy (SPDT). In the SPDT group, the cells were treated with sonication at an intensity of 0.5 W/cm2 and a frequency of 1 MHz, followed by different doses of light irradiation. The growth inhibition rate following treatment was determined by MTT assay. The apoptotic rate was examined by a flow cytometry. Cleavage of caspase 3, 8 and 9 was investigated by immunoblotting. Reactive oxygen species (ROS) were measured by a fluorescence microplate reader. The effect of SPDT on the glioma cells was also studied in the absence or presence of various ROS scavengers. The growth inhibition rate of C6 glioma cells treated with SPDT was significantly higher compared with sonodynamic therapy (SDT) or photodynamic therapy (PDT) alone at light doses <200 J/cm2. The growth inhibition rate of C6 glioma cells treated with SPDT did not rise significantly when the light dose increased to >120 J/cm2. The apoptosis rate was the highest in the SPDT group, when the light dose was at 80 J/cm2. A greater amount of ROS were generated in the SPDT group than in the groups treated with SDT or PDT alone. The addition of NaN3 or mannitol resulted in a decrease in the growth inhibition rate with SPDT. In conclusion, our data indicate that SPDT powerfully kills C6 glioma cells in vitro through the synergistic effects of SDT and PDT. The pathway of PDT inducing C6 glioma cell apoptosis includes both the mitochondrial and death receptor pathways. Furthermore, ROS may play an important role in SPDT.
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