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5-氨基乙酰丙酸介导的光动力疗法诱导HL-60细胞线粒体介导的凋亡 预览 被引量:1

Mitochondria-Mediated Apoptosis Induced by 5-Aminolevulinic Acid- Based Photodynamic Therapy in HL-60 Cells
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摘要 本研究探讨5-氨基乙酰丙酸介导的光动力疗法(ALA-PDT)作用于白血病细胞株HL-60后线粒体膜电位改变及凋亡相关基因Bcl-2表达的变化。以白血病细胞株HL-60为实验模型,实验分为4组:对照组、单纯ALA组、单纯光照组及ALA+PDT组。采用阳离子脂质荧光探针JC-1检测线粒体跨膜电位,RT-PCR及实时定量PCR方法检测PDT前后Bcl-2基因表达变化。结果表明,ALA-PDT后线粒体膜电位出现快速下降,0 h时线粒体跨膜电位崩塌的细胞比例升高至(58.28±0.92)%,与对照组相比有显著差异(P〈0.05)。随着时间延长,这种细胞比例逐步增多,而单纯ALA组和单纯光照组则无明显变化;ALA-PDT后2 h Bcl-2基因表达显著下降,4 h进一步下降达到谷底,24 h时仍呈低表达,并通过实时定量PCR进一步得到了证实。结论:ALA-PDT诱导的HL-60细胞凋亡过程与其影响线粒体跨膜电位有关,即通过影响线粒体功能促进细胞凋亡;ALA介导的光动力作用部分是通过在基因转录水平下调抗凋亡基因Bcl-2而促进凋亡的。 This study was purposed to investigate the changes of mitochondrial membrane potential (MMP) and apoptosis-related gene Bcl-2 expression of HL-60 cells treated with 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT). HL-60 cell line was used as a model and divided into 4 groups: ALA group, PDT group, ALA + PDT group and control group. The change of MMP was detected by flow cytometry with JC-1 (lipophilic cation 5,5' ,6,6'- tetrachloro-1,1',3,3 '-tetraethyl-benzimidazol-carbocyanine iodide) ; the mRNA expression of Bcl-2 was determined by semi-quantitative RT-PCR and real-time PCR. The results demonstrated that MMP significantly decreased after treatment with ALA-PDT and the ratio of cells with disrupted MMP obviously increased in ALA + PDT group in time-dependence manner, as compared with control, ALA and PDT groups ( P 〈 0.05 ), while no difference between ALA and PDT groups was found. The semi-quantitative RT-PCR and real-time PCR showed that the expression level of Bcl-2 was obviously down regulated at 2 h after ALA-PCT, further down-regulated at 4 h, and lasted in low level at 24 h. It is concluded that ALA-PDT-induced apoptosis of HL-60 ceils is associated with its effect on MMP, that is ALA-PDT promotes cell apoptosis through effect on mitochondrial function.
作者 韩晓凤 陈芳源 钟济华 欧阳仁荣 HAN Xiao-Feng, CHEN Fang-Yuan ,ZHONG Ji-Hua,OUYANG Ren-Rong Department of Hematology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 00, China Corresponding Author: CHEN Fang-Yuan, Senior Physician. Tel: (021)68383141. E-mail: chenfyO4@yahoo, com. cn
出处 《中国实验血液学杂志》 CAS CSCD 北大核心 2012年第1期 73-77,共5页 Journal of Experimental Hematology
关键词 5-氨基乙酰丙酸 光动力学治疗 HL-60细胞 细胞凋亡 线粒体 BCL-2 5-aminolevulinic acid photodynamic therapy HL-60 cell apoptosis mitochondrial Bcl-2
作者简介 陈芳源,主任医师.电话:(021)68383141.E-mail:chenfy04@yahoo.com.cn
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