利用基因表达谱芯片技术研究ALA-PDT对白血病细胞株HL-60的作用机制  

Mechanism Research of ALA-PDT in HL-60 Cells with DNA Microarray

作　　者：韩晓凤[1] 倪蓓文[1] 钟济华[1] 陈芳源[1] 

机构地区：[1]上海交通大学医学院附属仁济医院血液科,上海200127

出　　处：《应用激光》2016年第2期233-238,共6页Applied Laser

摘　　要：目的:研究探讨5-氨基乙酰丙酸介导的光动力疗法(ALA-PDT)对白血病细胞株HL-60的作用机制。方法:以白血病细胞株HL-60为实验模型。实验分为2组,对照组(未处理组)及ALA+PDT组。利用人全基因组基因表达谱芯片技术研究ALA-PDT后HL-60细胞基因改变情况。结果:人全基因组芯片共检测了48 000个基因,用RT-PCR的方法验证了数据的可靠性,对芯片结果产生的数据分析发现:ALA-PDT影响了细胞的多种生命进程,如转录调控,抑制翻译过程;促进一系列应激反应;上调促凋亡基因的表达,抑制抗凋亡基因的表达,促进凋亡的发生。结论:ALA-PDT通过诱导凋亡的方式杀伤HL-60细胞。多种调控机制参与凋亡的发生:c-Abl和PML基因是上调基因网络中的主导基因,共同参与了凋亡的发生。线粒体途径和TNF途径激活,DEDD2及CDC2L2介导的凋亡通路也占了一定的地位。ERN1、Bcl2、及c-Jun等基因参与其中促进凋亡的发生。Objective：To study the mechanisms of ALA-PDT-induced cell death in HL-60 leukemia cell line.Methods：Using HL-60 cell line as a model,the cells were divided into two groups,ALA＋PDT group and control group（no treatment）.Oligonucleotide microarray was used to detect the changes of gene expression profiles after ALA-PDT.Results：48,000 genes were involved in DNA microarrays.We evaluated the reliability of microarray platform using RT-PCR.Based on the results,many changes in transcription levels of the two cell lines treated by ALA-PDT occur：transcription regulation was activated;general translation was attenuated;genes involved in stress were upregulated;proapoptotic genes were upregulated and anti-apoptotic genes were downregulated.Conclusions：c-Abl and PML are the key genes in the network of upregulated genes.They both played an important role in apoptosis of HL-60 cells.The mitochondria pathway and TNF pathway were activated in HL-60 cells,followed by DEDD2 mediated caspase-independent pathway and CDC2L2 mediated caspase-dependent intrinsic pathway.And genes related to apoptosis such as ERN1,Bcl2 and C-Jun were significantly regulated,suggesting the involvement of multi-pathways during the process of ALA-PDT induced apoptosis.

关 键 词：5-氨基乙酰丙酸 光动力学治疗 HL-60 细胞凋亡 基因表达谱芯片技术 

分 类 号：R733.7[医药卫生—肿瘤]