新型水溶性卟啉类光敏剂A1光动力治疗黑色素瘤的实验研究

发布时间：2018-11-09 19:55

【摘要】：目的探讨新型水溶性卟啉类光敏剂A1光动力疗法对人黑色素瘤细胞系A375的体外及体内抑瘤效应及其相关机制,为临床应用提供理论依据。方法本研究分为空白对照组、单纯激光照射组、单纯光敏剂组及不同剂量光敏剂的光动力治疗组。MTT法检测光动力疗法对A375细胞增殖的影响。利用激光共聚焦显微镜检测A1在A375细胞中的亚细胞定位。利用Hoechst 33342染色和流式细胞术检测细胞凋亡及坏死情况。检测A1光动力治疗后,A375细胞中活性氧(reactive oxygen species,ROS)含量变化。建立黑色素瘤荷瘤小鼠模型,绘制肿瘤生长曲线,观察治疗效果。结果体外实验表明,单纯给药或光照对肿瘤细胞的生长无或很小影响,而A1-PDT治疗组能明显抑制A375肿瘤细胞增殖。Hoechst 33342染色和流式细胞术结果显示,与对照组比较,A1-PDT治疗组可以明显诱导A375细胞凋亡。与对照组比较,A1-PDT治疗组可以明显增加A375细胞中ROS含量。激光共聚焦显微镜显示化合物A1主要定位在肿瘤细胞的溶酶体中。体内实验显示,光动力疗法可以明显抑制小鼠黑色素瘤皮下移植瘤生长。结论光敏剂A1介导的光动力疗法对黑色素瘤细胞及移植瘤的生长抑制作用明显。光动力疗法以细胞凋亡为主,其机制可能与光敏剂A1定位在肿瘤细胞的溶酶体,增加A375细胞中ROS含量,降低谷胱甘肽(GSH)含量相关。
[Abstract]:Objective to investigate the antitumor effect of a new water-soluble porphyrin type Guang Min A1 on human melanoma cell line A375 in vitro and in vivo and to provide theoretical basis for clinical application. Methods the study was divided into three groups: blank control group, laser irradiation group, simple Guang Min group and different dosage of Guang Min group. The effect of photodynamic therapy on the proliferation of A375 cells was detected by MTT method. The subcellular localization of A1 in A375 cells was detected by laser confocal microscopy. Apoptosis and necrosis were detected by Hoechst 33342 staining and flow cytometry. The content of reactive oxygen species (reactive oxygen species,ROS) in A375 cells was measured after A 1 photodynamic therapy. The model of melanoma bearing mice was established, the tumor growth curve was drawn and the therapeutic effect was observed. Results in vitro experiments showed that there was no or little effect on the growth of tumor cells in A1-PDT group, while the proliferation of A375 tumor cells was significantly inhibited in A1-PDT treatment group. The results of Hoechst 33342 staining and flow cytometry showed that, compared with the control group, the growth of A375 tumor cells was significantly inhibited. Apoptosis of A375 cells was induced in A1-PDT group. Compared with the control group, the ROS content in A375 cells was significantly increased in A1-PDT treatment group. Laser confocal microscopy showed that compound A1 was mainly localized in lysosomes of tumor cells. In vivo experiments showed that photodynamic therapy could significantly inhibit the growth of murine melanoma subcutaneous xenografts. Conclusion Guang Min A 1-mediated photodynamic therapy can inhibit the growth of melanoma cells and xenografts. The mechanism of photodynamic therapy is mainly apoptosis, which may be related to the localization of Guang Min A1 in lysosomes of tumor cells, the increase of ROS content and the decrease of glutathione (GSH) content in A375 cells.
【作者单位】：武警后勤学院救援医学系生药学教研室;中国医学科学院生物医学工程研究所;
【基金】：武警后勤学院博士启动金(No WHTD201304-2)
【分类号】：R739.5

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