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目的:探讨光敏剂焦脱镁叶绿酸-a甲酯(pyropheophorbide-a methyl ester,MPPa)介导的光动力治疗(photodynamic therapy,PDT)对顺铂(cisplatin,DDP)耐药的肺癌细胞的作用。方法:DDP、PDT和PDT+DDP处理肺癌A549和A549/DDP细胞后,应用CCK-8法、FCM法、2’,7’-二氯荧光黄双乙酸盐(2’,7’-dichlorodihydrofluorescein diacetate,DCFH-DA)和蛋白质印迹法分别检测细胞增殖、细胞凋亡、细胞内活性氧(reactive oxygen species,ROS)水平及caspase-3、Bcl-2和Bax蛋白表达水平。结果:PDT组A549和A549/DDP细胞增殖率均低于对照组(A549和A549/DDP细胞未进行任何处理)(P值均<0.05),PDT+DDP组A549和A549/DDP细胞增殖率均低于对照组、DDP组和PDT组(P值均<0.05)。PDT+DDP作用A549和A549/DDP细胞的联合指数分别为1.11和1.10,具有相加效应。PDT+DDP组A549和A549/DDP细胞凋亡率和ROS水平均高于对照组、DDP组和PDT组(P值均<0.05)。PDT组和PDT+DDP组A549和A549/DDP细胞中caspase-3和Bax蛋白表达水平均高于对照组,而Bcl-2蛋白表达水平均低于对照组(P值均<0.05)。结论:PDT或联合DDP对肺癌A549/DDP细胞具有明显杀伤作用,主要通过ROS.线粒体途径促进细胞凋亡;PDT与DDP具有相加效应。
Objective: To investigate the effect of photodynamic therapy (PDT) mediated by photosensitizer pyropheophorbide-a methyl ester (MPPa) on cisplatin (DDP) -resistant lung cancer cells effect. Methods: After treated with DDP, PDT and PDT + DDP, the cell lines of A549 and A549 / DDP were treated with CCK-8, FCM, 2’7’-dichlorodihydrofluorescein diacetate DCFH-DA) and Western blotting were used to detect the cell proliferation, apoptosis, the level of reactive oxygen species (ROS) and the protein expression of caspase-3, Bcl-2 and Bax. Results: The proliferation rates of A549 and A549 / DDP cells in PDT group were significantly lower than those in control group (P <0.05). The proliferation rates of A549 and A549 / DDP cells in PDT + DDP group were significantly higher than those in control group Lower than the control group, DDP group and PDT group (P <0.05). PDT + DDP effect The combined index of A549 and A549 / DDP cells were 1.11 and 1.10, respectively, with an additive effect. The apoptosis rate and ROS level of A549 and A549 / DDP cells in PDT + DDP group were higher than those in control group, DDP group and PDT group (P <0.05). The protein expressions of caspase-3 and Bax in PDT group and PDT + DDP group were higher than those in control group, while the expression levels of Bcl-2 protein in PDT + DDP group were lower than those in control group (all P <0.05). CONCLUSION: PDT or DDP can significantly kill A549 / DDP cells and promote cell apoptosis mainly through ROS mitochondria pathway. PDT and DDP have additive effects.