【摘要】 目的 :用反义 bcr/abl寡核苷酸片段诱导 K562细胞凋亡 ,观察 Caspase3表达及活性变化 ,以阐明 Caspase3在慢性髓性白血病 ( CML)发病机制中的作用。方法 :用反义 bcr/abl寡核苷酸片段 AS和对照无义片段 NS处理 CML急变细胞系 K562 ,检测细胞凋亡及 Caspase3表达和活性的变化。结果 :K562细胞经 AS处理 72 h,流式细胞仪检测未见亚二倍体细胞增多。原位末端标记法显示 AS组 2 4 h始出现细胞凋亡增多 ,72 h达高峰 ,为 ( 78.50± 6.60 ) % ,较对照组 ( 1 8.2 1± 3.32 ) %及 NS组 ( 2 4 .1 3± 4.1 7) %明显增多 ( P<0 .0 5)。Western blot检测显示 ,培养 72 h AS组 Caspase3原酶及 p17活性亚单位的表达增强。荧光分光光度法测定 Caspase3活性示 ,AS处理 1 2 h Caspase3活性即已升高 ,48h达高峰 ,为处理前的 4.71倍。对照组及NS组上升幅度较小 ,72 h最高 ,分别为处理前的 2 .78倍和 2 .30倍。结论 :Caspase3参与了 CML的 bcr/abl基因抑制细胞凋亡的作用。更多还原

【Abstract】 Objective: To explore the role of Caspase 3 in the pathogenesis of bcr/abl gene inhibiting apoptosis in CML. Methods: CML blast crisis cell line K562 cells were incubated with bcr/abl antisense oligomer(AS) and the control nonsense oligomer(NS). Apoptosis and Caspase3 expression and activity were detected. Results: Apoptosis of K562 exposed to AS for 72h was significantly increased than in the control and NS groups when detected by ISEL(78.50±6.60)% while not by cytometry. Caspase3 proenzyme and its p 17 subunit were also increased when detected by Western Blot. Increasement of Caspase3 activity detected by fluorometry could be seen when the K562 cells were exposed to AS for 12h, and reached the peak at 48h, which was 4.71 times higher than that before the treatment. Compared with the AS group the Caspase3 activity of control and NS groups was much lower reaching their highest levels at 72h and being 2.78 and 2.30 times respectively than pretreatment. Conclusion: Caspase3 plays a role in the pathogenesis of bcr/abl gene inhibiting apoptosis in CML and may be an early event of apoptosis.更多还原