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目的探讨藻蓝蛋白介导的光动力疗法治疗小鼠HeLa细胞瘤的免疫和凋亡机制。方法将HeLa细胞接种于小鼠肋缘皮下脾区构建荷瘤小鼠模型。实验分成3组:对照组、激光照射组、光动力治疗组(PDT组)。PDT组:肿瘤局部皮下注射藻蓝蛋白液0.25ml(约2g)2h后以He-Ne激光照射。实验第7d测瘤块重量,取胸腺、脾脏检测NK细胞活性和T细胞增殖活性。取瘤块制成石蜡包埋切片,采用原位核酸杂交技术、免疫组织化学技术检测肿瘤细胞内CD44、P53、NFκB、Fas基因的表达。结果与对照组相比,激光照射组NK细胞和免疫T细胞的增殖活性有所增强,肿瘤细胞内抗凋亡基因(Fas)表达量显著增多,而瘤块的重量、肿瘤形成率和抗凋亡基因(P53、NFκB、CD44)明显减少。以上各项指标PDT组与激光组比较,差异亦具有显著或非常显著意义(P>0.05或P<0.01)。结论藻蓝蛋白介导的光动力疗法通过增强机体的免疫力同时启动HeLa细胞内的凋亡信号转导通路诱导细胞凋亡,从而达到杀死肿瘤的目的。
Objective To investigate the mechanisms of photoprotection mediated by phycocyanin in the treatment of mouse HeLa cell tumor. Methods HeLa cells were inoculated into the subcutaneous splenic region of the cosmid in mice to construct a tumor-bearing mouse model. The experiment was divided into three groups: control group, laser irradiation group and PDT group. PDT group: the tumor subcutaneous injection of phycocyanin 0.25ml (about 2g) 2h He-Ne laser irradiation. On the 7th day, the weight of the tumor was measured, and the thymus and spleen were used to detect the activity of NK cells and T cell proliferation. Tumor specimens were made into paraffin embedded sections and the expression of CD44, P53, NFκB and Fas genes in tumor cells were detected by in situ hybridization and immunohistochemistry. Results Compared with the control group, the proliferative activity of NK cells and immune T cells in laser irradiation group was enhanced, and the expression of anti-apoptotic gene (Fas) in tumor cells was significantly increased. The weight, tumor formation rate and anti- The death genes (P53, NFκB, CD44) were significantly reduced. The above indicators PDT group compared with the laser group, the difference also has significant or very significant (P> 0.05 or P <0.01). Conclusions Phycoerythrin-mediated photodynamic therapy can induce apoptosis in HeLa cells by increasing the immunity of HeLa cells, and thus killing the tumor.