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 study the treatment of Hela tumor in mice with C-phycocyanin(C-PC) mediated photodynamic therapy(PDT) and its mechanism.Methods HeLa lines were injected into spleen area of mice to establish mice tumor models which then were divided into control group,He-Ne laser radiation group and PDT group.7 days later,tumor weight,activities of NK cells,proliferation activities of immunocytes were determined.Tumor tissue was cut into sections and the expressions of apoptosis-related genes such as CD44,P53,NfκB and Fas in HeLa cells in vivo were determined by in situ hybridization assays and immunohistochemistry.ResultsCompared with control group,immunocyte proliferation ability and pro-apoptotic gene expression in He-Ne radiation group were slighthly higher,while the tumor weight and forming rate,anti-apoptotic genes quantities were comparatively lower.When the mice were treated with C-PC followed by He-Ne laser radiation,the differences were much greater.ConclusionsC-PC is confirmed to be one kind of photosensitizer,and when HeLa cells are treated with C-PC prior to He-Ne laser radiation,the antitumor activities are much stronger and can enhance immunity and faciliate the transduction of tumor apoptosis signals.