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目的:通过细胞学水平研究不同剂量ALA光动力学治疗对C6胶质瘤细胞细胞凋亡的影响。方法:以不同剂量的ALA(10μg/ml、20μg/ml、40μg/ml、80μg/ml、160μg/ml)与C6胶质瘤细胞一起培养,随后以630nm半导体激光200mW/cm2,照射肿瘤细胞,照射20分钟,及与不用光敏剂单照激光、单用光敏剂不照光、不用光敏剂不照激光空白对照组比较。以流式细胞仪及倒置微镜、电镜观察促使肿瘤细胞凋亡或死亡的差别,寻找促使肿瘤细胞凋亡或死亡的最小合适剂量。结果:对照组没有细胞凋亡,ALA-PDT组随着剂量的增加细胞凋亡数增加。尤其晚期凋亡数明显增加,用药剂量增加至40μg/cc才能达到活细胞少于50%,即使剂量增加到160μg/cc,还是有19.7423的活细胞存在。结论:单纯用ALA激光PDT治疗不能达到完全将肿瘤细胞杀灭,ALA40mg/kg激光光动力学治疗才能得到明显的治疗效果。
OBJECTIVE: To study the effects of different dosages of ALA photodynamic therapy on C6 glioma cell apoptosis by cytology. Methods: C6 glioma cells were cultured with different doses of ALA (10μg / ml, 20μg / ml, 40μg / ml, 80μg / ml, 160μg / ml), then irradiated with 630nm semiconductor laser at 200mW / cm2, Irradiation for 20 minutes, and with no photosensitizer single-shot laser, single-use photosensitizer does not light, no photosensitizer does not light the blank control group comparison. Using flow cytometry and inverted micromirror, electron microscopy to promote the difference of apoptosis or death of tumor cells, looking for the smallest suitable dose to promote tumor cell apoptosis or death. Results: There was no apoptosis in the control group. The number of apoptotic cells increased with the increase of dose in ALA-PDT group. In particular, the number of late apoptotic cells increased significantly, and the dosage increased to 40μg / cc to reach less than 50% of the viable cells. Even though the dose increased to 160μg / cc, 19.7423 live cells were still present. Conclusion: ALT laser PDT alone can not achieve the complete killing of tumor cells, ALA 40mg / kg laser photodynamic therapy in order to get a significant therapeutic effect.