三氧化二砷联合硼替佐米对K562细胞增殖和凋亡的影响及其机制的实验研究  

Effects of Arsenic Trioxide Combined with Bortezomib on Proliferation and Apoptosis of K562 Cells and Their Mechanism

作　　者：吴弘英[1] 王松梅[1] 黄俊谦[1] 郝鲁梅[1] 李雪梅[1] 卫丽妍[1] 

机构地区：[1]青岛市市立医院血液科

出　　处：《中国实验血液学杂志》2012年第6期1361-1364,共4页Journal of Experimental Hematology

摘　　要：本研究探讨三氧化二砷(As2O3)单独或联合硼替佐米(Bor)对髓系白血病细胞系K562细胞增殖、凋亡的影响及其可能的作用机制。不同浓度的As2O3、Bor处理细胞24、48 h,采用MTT比色法检测细胞增殖活性,流式细胞术检测单用As2O3、Bor及二者联用的细胞凋亡率及细胞周期的变化;免疫组织化学法分析各组核转录因子(NF-κB)的表达水平。结果表明,As2O3(0.1-0.8μmol/L)和Bor(5-50 nmol/L)均能抑制K562细胞的增殖,且与作用时间和药物浓度有关,时间越长,浓度越大,抑制越明显(P<0.05);Bor和As2O3作用48 h的IC50值分别为20nmol/L和0.6μmol/L。As2O3(0.5μmol/L)或Bor(10 nmol/L)分别单独处理细胞24 h,K562细胞凋亡增加,联合用药组的细胞凋亡率高于单药组(P<0.01)。As2O3或Bor处理K562细胞6 h时细胞周期分布显示:As2O3组细胞阻滞于G0/G1期,Bor组细胞阻滞于G2/M期。As2O3或Bor分别单独处理细胞24 h,NF-κB表达阳性率及积分较对照组明显降低(P<0.05),联合用药组的NF-κB表达阳性率及积分较As2O3及Bor单用组均明显降低(P<0.01)。结论:As2O3及Bor单药均可以直接抑制K562细胞增殖,促进凋亡,两药小剂量联合作用后,细胞增殖抑制及促凋亡作用较单药明显增强。细胞周期阻滞点的不同及NF-κB表达的下降可能是As2O3和Bor联用后协同作用的机制之一。This study was aimed to investigate the effect of arsenic trioxide（ As2O3 ） alone and in combination with bortezomib（Bor） on proliferation and apoptosis of leukemia cell line K562, and to analyze the potential mechanism. K562 cells were treated with different concentrations of As2O3 or Bur （ alone or combination） for 24, 48 h. MTT method was used to detect the cell proliferation. After K562 cells were treated with 0.5 μmol/L As2O3 alone or in combination with 10 nmoL/L Bor, the apoptosis rate and cell cycle were measured by flow cytometry, and the activity of NF-κB was analyzed by SP immunohistochemistry. The results indicated that the different concentrations of AS203 and Bor could inhibit the K562 cell proliferation in a time- and dose-dependent manners （P 〈 0.05）. The IC50 of Bor and As2O3 in 48 h were 20 nmol/L and 0.6 μmol/L respectively. When K562 cells were treated with As203 or Bor alone for 24 h, the apoptotic rate of K562 cells increased, and the apoptotic rate in combination group was higher than that in As203 or Bor group. The cells were apparently arrested in G2/M phase in Bor group and G0/G1 phase in AS2O3 group. The activity of NF-κB decreased significantly in As2O3 or Bor group （P 〈 0.05 ）, this effect was most significant in the combination group （P 〈0.01 ）. It is concluded that both As2O3 and Bor can inhibit the proliferation and induce apoptosis of K562 cells, a synergistic effect can be observed when a low dose of As2O3 combined with low dose of Bor. The different cell cycle block site and the decrease of activity of NF-κB may be one of the mechanisms underlying their synergic effect.

关 键 词：三氧化二砷 硼替佐米 K562细胞 细胞凋亡 细胞周期 核因子-ΚB 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]