论文部分内容阅读
目的:本研究观察了5-氨基乙酰丙酸介导的光动力治疗(ALA-PDT)对耐药白血病细胞株HL-60/ADR的杀伤作用及对耐药白血病原代细胞存活率的影响。方法:以耐药白血病细胞株HL-60/ADR为实验模型,同时在11例白血病患者原代细胞中进行检测。实验分为4组,对照组、单纯ALA组、单纯光照组及ALA+PDT组。用MTT法测定细胞的存活率,采用阳离子脂质荧光探针JC-1检测线粒体跨膜电位,用Real-time PCR检测PDT前后HL-60/ADR细胞株中bcl-2基因及多药耐药基因MRP的表达变化。结果:ALA-PDT后HL-60/ADR细胞株线粒体跨膜电位出现快速下降,0,2,4h时线粒体跨膜电位崩塌的细胞比例分别升高至55.91%±2.60%、64.27%±1.08%、82.17%±0.43%,与对照组相比皆有显著差异(P<0.05),呈时间依赖性。而单纯ALA组和单纯光照组则无明显变化。HL-60/ADR细胞株在ALA-PDT后Bcl-2和MRP基因均呈明显下降趋势。在初发和复发难治急性白血病原代细胞中ALA+PDT组均显示较强的光动力效应。结论:ALA-PDT诱导的HL-60/ADR细胞的杀伤可能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。同时表明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而促进凋亡的发生,另一方面通过下调耐药基因MRP的表达而部分逆转耐药。同时ALA-PDT对原代白血病细胞同样有较大的抑制作用。
OBJECTIVE: We studied the effects of 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) on the killing of drug-resistant leukemia cell line HL-60 / ADR and its effect on the survival of primary drug-resistant leukemia cells. Methods: The experimental HL-60 / ADR cell line was used as an experimental model, and 11 primary leukemia patients were also detected. The experiment was divided into 4 groups, control group, simple ALA group, simple light group and ALA + PDT group. Cell viability was measured by MTT assay. Mitochondrial transmembrane potential was measured by cationic lipid fluorescent probe JC-1. The bcl-2 gene and multidrug resistance in HL-60 / ADR cell lines were detected by Real-time PCR Gene MRP expression changes. Results: After ALA-PDT, the mitochondrial transmembrane potential of HL-60 / ADR cells rapidly decreased. The percentage of cells with mitochondrial transmembrane potential collapse at 0, 2 and 4 h increased to 55.91% ± 2.60% and 64.27% ± 1.08% , 82.17% ± 0.43%, which were significantly different from the control group (P <0.05) in a time-dependent manner. The simple ALA group and simple light group no significant change. HL-60 / ADR cell lines in ALA-PDT Bcl-2 and MRP gene showed a significant downward trend. The ALA + PDT group showed a strong photodynamic effect in primary and relapsed refractory acute leukemia primary cells. CONCLUSION: ALA-PDT-induced killing of HL-60 / ADR cells may be related to its effect on mitochondrial transmembrane potential, that is, mitochondrial function is promoted through apoptosis. At the same time, ALA-mediated photodynamic therapy partly promoted apoptosis by down-regulating the anti-apoptotic gene Bcl-2 at the gene transcription level and partially reversed the resistance by down-regulating the expression of the MRP gene. At the same ALA-PDT on the same primary leukemia cells have a greater inhibitory effect.