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目的探讨高选择性的蛋白酶抑制药 MG132诱导的 K562细胞凋亡及其对转录核因子B(NF-B)活性和抗凋亡蛋白生存素(Survivin)表达的影响。方法采用细胞形态学观察和流式细胞术检测细胞凋亡,免疫细胞化学和 Western 印迹分析 NF-B 的活性和 Survivin 的表达。结果不同浓度的 MG132作用24 h 后可诱导 K562细胞凋亡,呈剂量依赖效应。NF-B 和 Survivin 在 K562细胞中异常高表达,MG132作用后均明显下调。2、4、6、8μmol/L MG132作用于 K562细胞24 h 后,NF-B 活性分别下凋至作用前的75.0%±3.7%、59.9%±5.3%、45.4%±5.7%和25.0%±4.2%,而Survivin 蛋白表达分别下调至作用前的90.9%±10.1%、66.7%±5.2%、45.4%±5.7%和30.3%±6.6%,NF-κB活性降低的同时 Survivin 蛋白的表达也下调,两者密切相关(Pearson 相关系数0.989,P<0.01)。结论 MG132可有效地抑制 NF-κB的活性,诱导 K562细胞发生凋亡,同时抗凋亡蛋白Survivin 的表达也随之下调。
Objective To investigate the apoptosis of K562 cells induced by highly selective protease inhibitor MG132 and its effect on the expression of nuclear factor-B (NF-B) and anti-apoptotic protein Survivin. Methods Cell apoptosis was detected by morphological observation and flow cytometry. The activity of NF-B and the expression of Survivin were analyzed by immunocytochemistry and Western blotting. Results Different concentrations of MG132 induced apoptosis of K562 cells in a dose-dependent manner. NF-B and Survivin in K562 cells were abnormally high expression, MG132 significantly decreased after treatment. After treated with 2,4,6,8μmol / L MG132 for 24 h, NF-|ÊB activity reached 75.0% ± 3.7%, 59.9% ± 5.3%, 45.4% ± 5.7% and 25.0% ± 4.2%, while the expression of Survivin protein was down-regulated to 90.9% ± 10.1%, 66.7% ± 5.2%, 45.4% ± 5.7% and 30.3% ± 6.6% , The two are closely related (Pearson correlation coefficient 0.989, P <0.01). Conclusion MG132 can effectively inhibit the activity of NF-κB and induce the apoptosis of K562 cells. Meanwhile, the expression of anti-apoptotic protein Survivin is also down-regulated.