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目的:研究喜树碱(CPT)诱导的人早幼粒细胞性白血病细胞HL-60凋亡过程中线粒体的质量和膜电势的变化。方法:以CPT诱导HL-60细胞凋亡为模型,利用膜联蛋白V(annexinV)-FITC/PI双染流式细胞术,研究HL-60细胞的凋亡和坏死。用DiOC6(3)染色流式细胞术,检测线粒体的膜电势(△ψm)。用NAO染色流式细胞术,检测线粒体的质量。结果:在4×10-6mol/LCPT的诱导下,HL-60细胞(12h)早期的凋亡率为(12.75±4.61)%,对照组为(2.88±2.49)%,二者相比较差异显著(P<0.01);CPT组坏死比率为(3.48±1.67)%,对照组为(0.71±1.10)%(P<0.01)。PI染色的结果显示,HL-60细胞(12h)晚期凋亡细胞的百分率,CPT组为(3.52±1.07)%,对照组为(0.46±1.06)%(P<0.01)。同时观察到,G2/M期细胞出现阻滞,G2/M期细胞的百分率,对照组为(22.46±2.19)%,CPT组为(13.45±1.91)%(P<0.01)。在12h时间点,CPT组线粒体的质量显著低于对照组(P<0.01),低线粒体质量的细胞所占百分率,对照组为(4.53±1.26)%,CPT组为(25.74±2.09)%。同时,CPT组线粒体的膜电势显著下降(P<0.01),CPT组线粒体膜电势降低的比率为(17.71±5.23)%,对照组为(1.64±2.00)%。结论:CPT诱导HL-60细胞凋亡过程中,线粒体的质量和膜电势均有所下降,但其去极化作用增强。
OBJECTIVE: To study the changes of mitochondrial mass and membrane potential during the apoptosis of human promyelocytic leukemia HL-60 cells induced by CPT. Methods: The apoptosis of HL-60 cells was induced by CPT. The apoptosis and necrosis of HL-60 cells were studied by annexin V-FITC / PI double staining. Mitochondrial membrane potential (△ ψm) was measured by DiOC6 (3) staining. Mitochondrial mass was detected by NAO staining and flow cytometry. Results: The apoptotic rates of HL-60 cells (12.75 ± 4.61)% and (2.88 ± 2.49)%, respectively, in HL-60 cells induced by 4 × 10-6 mol / L CPT were significantly different (P <0.01). The necrosis rate was (3.48 ± 1.67)% in CPT group and (0.71 ± 1.10)% in control group (P <0.01). PI staining showed that the percentage of apoptotic cells in late HL-60 cells (12h) was (3.52 ± 1.07)% in CPT group and (0.46 ± 1.06)% in control group (P <0.01). At the same time, the percentage of cells in G2 / M phase was arrested in G2 / M phase, which was (22.46 ± 2.19)% in control group and (13.45 ± 1.91)% in CPT group (P <0.01). At 12h, the mitochondrial mass in CPT group was significantly lower than that in control group (P <0.01). The percentage of cells with low mitochondrial mass was (4.53 ± 1.26)% in control group and (25.74 ± 2.09)% in CPT group. At the same time, the membrane potential of mitochondria in CPT group decreased significantly (P <0.01), the mitochondrial membrane potential decreased (17.71 ± 5.23)% in CPT group and (1.64 ± 2.00)% in CPT group. CONCLUSION: During CPT-induced HL-60 cell apoptosis, the mitochondrial mass and membrane potential decrease, but the depolarization effect is enhanced.