摘要
目的探讨藻蓝蛋白介导的光动力学疗法在乳腺癌治疗中的机制。方法将MCF-7细胞接种于小鼠肋缘皮下脾区构建乳腺癌小鼠模型。小鼠分成4组:对照组、He-Ne激光照射组、藻蓝蛋白处理组、光动力学治疗组(PDT组)。10d后检测瘤块重量,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组织细胞内凋亡相关蛋白的表达。体外培养的MCF-7细胞也相应分成4组,通过MTT法、电镜和免疫荧光技术检测细胞增殖活性、细胞形态、细胞色素C表达量的变化。结果与对照组相比,激光照射组各检测指标均无明显差异,而藻蓝蛋白处理组中NK细胞和T细胞的增殖活性有所增强,肿瘤组织细胞内抗凋亡蛋白(Fas)表达量明显增多,而瘤块的重量、肿瘤形成率和抗凋亡蛋白(p53、NF-κB、CD44)明显减少,如果藻蓝蛋白结合激光治疗发现各检测指标与对照组比较差异更为明显。体外试验证实藻蓝蛋白能抑制MCF-7细胞的增殖,促进细胞色素C的释放,电镜下细胞呈现典型的凋亡形态,用光动力学方法处理后效果更为明显。结论藻蓝蛋白可以作为一种光敏剂,其介导的光动力学疗法通过增强机体的免疫力同时启动乳腺癌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。
Aim To study the mechanism of C-phycocyanin（C-PC） mediated photodynamic therapy（PDT） in curing breast cancer.Methods MCF-7 cells were injected into spleen area of mice to establish mice models with breast cancer which then were divided into four groups： control group,He-Ne laser radiation group,C-PC treatment group and PDT group.10 days later,tumor weight,activities of NK cells,proliferative activities of T cells were determined.The tumors were made to paraffin section and the expressions of apoptosis related proteins such as CD44,P53,NFκB and Fas in MCF-7 cells in vivo were determined by in situ hybridization assays and immunohistochemistry.MCF-7 cells were cultured in vitro and divided into four groups.Subsequently,MTT,electron microscope and immunofluorescence assay were applied to explore the changes of cellular proliferative activities,morphological features,and quantities of cytochrome c release in vitro MCF-7 cells.Results Compared with control group,He-Ne laser alone was not evidently effective on the items examined.However,in C-PC treated mice,NK cells activities,T cell proliferation ability and pro-apoptotic protein quantities were a bit higher,while the tumor weight and forming rate,anti-apoptotic protein quantities were comparatively lower in MCF-7 cells in tumor.When the mice were treated with C-PC followed by He-Ne laser radiation,the differences were much enlarged.The results on MCF-7 cells in vitro proved that C-PC was able to inhibit cell proliferation and cause ultrastructural changes.Moreover,cytochrome C release was enhanced after C-PC treatment.When MCF-7 cells were treated with PDT,the functional effects became significantly greater.Conclusions C-PC is confirmed to be one kind of photosensitizer.The mechanism responsible for the antitumor activities is likely that it can enhance immunity and faciliate the transduction of MCF-7 breast cell apoptosis signals.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第3期383-389,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No81001346)
山东省中青年科学家科研奖励基金资助项目(No2007BS03016)
山东省教育厅科学研究项目(NoJ07YE16-2)
