摘要
目的观察MPPa光动力作用对人鼻咽癌细胞生长的抑制作用及可能机制。方法采用倒置显微镜观察和MTT法检测MPPa光动力作用后人鼻咽癌细胞株CNE2细胞生长状况;应用膜联蛋白Annexin V-PI双染结合流式细胞仪分析和细胞核染色观察MPPa光动力作用后人鼻咽癌细胞株CNE2细胞凋亡的发生情况;罗丹明123结合流式细胞仪分析MPPa光动力作用后人鼻咽癌细胞株CNE2细胞线粒体膜电位的变化。结果MPPa光动力作用显著抑制人鼻咽癌细胞株CNE2细胞的生长,在低浓度MPPa和低能量密度光照条件下其抑制作用呈剂量效应关系,倒置显微镜下观察到MP-Pa光动力作用组癌细胞数量显著减少,细胞脱落、固缩;MPPa光动力作用人鼻咽癌细胞株CNE2细胞发生凋亡和继发性坏死的比率均显著高于单纯光照射组、单纯MPPa光敏剂处理组和假照射组(P<0.01),而单纯光照组、单纯MPPa光敏剂处理组和假照射组三对照组间无明显差异(P>0.05),Hoechst33258细胞核荧光染色荧光显微镜下观察到MPPa光动力作用后癌细胞核固缩、碎裂,可见凋亡小体。MPPa光动力作用人鼻咽癌细胞株CNE2细胞线粒体膜电位下降。结论MPPa光动力作用不仅能有效抑制诱导人鼻咽癌细胞株CNE2细胞生长,而且可诱导其发生凋亡,且MPPa光动力作用可能通过线粒体通路诱导鼻咽癌细胞凋亡。
Objective To nasopharyngeal carcinoma cells explore photodynamic inhibition of pyropheophorbide-a methyl ester （MPPa） in and the mechanism. Methods The poorly differentiated CNE2 cell line was chosen to investigate the photocytotoxic potency and efficacy of MPPa 24 h after PDT by MTT reduction assay. The apoptosis of CNE2 cells was analyzed 8 h after photosensitization of MPPa （2 μmol/L） under red light irradiation at a light dose of 2 J/cm^2 by/low cytometry with Annexin V-FITC and PI staining and nuclear staining with Hoechst 33258, and the changes in mitochondrial membrane potential （△ψm） were monitored by/low cytometry with Rhodamine123 staining. Results Drug- and light-dose response curves were observed, showing that MPPa mediates a dose- and light-dependent phototoxicity in the NPC/CNE2 cells. MPPa could significantly trigger the NPC/CNE2 cells into apoptosis and the collapse of △ψm were observed in the NPC/CNE2 cells treated by PDT with MPPa. Conclusion MPPa can effectively photokill the NPC/CNE2 cells and induce ap- optosis as a prominent mode of cell death probably through mitochondria-initiated apoptosis pathway.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第14期1457-1460,共4页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目(30300468)
全军医学科研"十五"计划青年基金资助项目(01Q102)
香港浸会大学FacultyResearchGrant(FRG)基金资助项目(FRG/03-04/L-17)
重庆市中医药管理局科学基金资助项目(渝中医[2004]12号)
重庆市卫生局科学基金资助项目(渝卫科技[2004]53号)~~
关键词
光动力治疗
MPPa
鼻咽癌细胞
细胞凋亡
线粒体膜电位
photodynamic therapy
MPPa
nasopharyngeal carcinoma
apoptosis
mitochondrial membrane potential
