应用激光, 2016, 36 (2): 233, 网络出版: 2016-06-06  

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
作者单位
上海交通大学医学院附属仁济医院血液科, 上海 200127
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。
Abstract
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.
参考文献

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韩晓凤, 倪蓓文, 钟济华, 陈芳源. 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[J]. 应用激光, 2016, 36(2): 233. Han Xiaofeng, Ni Beiwen, Zhong Jihua, Chen Fangyuan. Mechanism Research of ALA-PDT in HL-60 Cells with DNA Microarray[J]. APPLIED LASER, 2016, 36(2): 233.

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