As_2S_2与STI 571协同诱导K562细胞凋亡  

Synergism of As_2S_2 and STI 571 in inducing apoptosis of K562 cells

作　　者：李俊娥[1] 孙关林[1] 吴英理[1] 邬维礼[1] 

机构地区：[1]上海第二医科大学附属瑞金医院,上海血液学研究所,200025

出　　处：《中华血液学杂志》2003年第11期580-583,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金资助 (3 9470 3 18)

摘　　要：目的　探索As2 S2 和STI 5 71联合使用对K5 6 2细胞的诱导凋亡作用及其机制。方法　用细胞计数法研究As2 S2 对K5 6 2细胞的生长抑制作用 ;细胞凋亡的检测用流式细胞术、基因组DNA电泳、细胞形态学观察等方法 ;蛋白表达的检测采用Western blot法 ;基因表达的变化用半定量RT PCR法。结果　As2 S2 1～ 5 μmol/L作用 2 4～ 72h ,即可明显抑制K5 6 2细胞增殖 ,3～ 5 μmol/L作用 4 8～ 72h即可诱导K5 6 2细胞凋亡。 3μmol/LAs2 S2 作用 72h ,细胞凋亡率达 34.4 % ;5 μmol/LAs2 S2 作用 4 8,72h ,细胞凋亡率分别达 2 1.8%和 4 6 .0 %。 1μmol/LSTI 5 71作用 4 8h ,细胞凋亡率为 (18.4± 1.4 ) % ;5 μmol/LAs2 S2 作用 4 8h ,细胞凋亡率为 (15 .8± 1.2 ) % ;而两者合用细胞凋亡率上升为 (40 .6± 2 .0 ) %。对于U937细胞 ,单用 1μmol/LSTI5 71作用 4 8h ,细胞凋亡率为 (4.5± 1.1) % ;单用 5 μmol/LAs2 S2 作用 4 8h ,细胞凋亡率为 (6 .0± 1.2 ) % ;而两者合用 ,细胞凋亡率为 (7.3± 1.0 ) % ,无明显协同作用。As2 S2 能降低K5 6 2细胞中Bcr Abl蛋白水平 ,并抑制c Abl和Bcr AblPTK活性 ,但As2 S2 并不调变bcr abl基因表达水平。结论　As2 S2 联合STI 5 71可增强诱导K5 6 2细胞凋亡的作用 。Objective To investigate the synergistic effect of As_2S_2 and STI 571 on K562 cells and its mechanism. Methods The inhibitive effect of As_2S_2 on the proliferation of K562 cells was determined by cell number count.Cell apoptosis was assessed by flow cytometry, DNA fragmentation and morphology. Protein expression was determined by Western-blot and gene expression by RT-PCR. Results As_2S_2 could significantly inhibit the proliferation and induce apoptosis of K562 cells in a dose and time-dependent manner at concentrations from 1 μmol/L to 5 μmol/L for 24～72 h. 34.4%, 21.8% and 46.0% of the treated-cells displayed apoptosis at 3 μmol/L for 72 h, 5 μmol/L for 48 h and 5 μmol/L for 72 h, respectively. Compared to treatment with STI571 (0.25～1.00 μmol/L) or As_2S_2 (1～5 μmol/L) alone, treatment of K562 cells with As_2S_2 and STI571 combination induced more cell apoptosis. (18.4±1.4)% and (15.8±1.2)% cells underwent apoptosis at 1 μmol/L STI571 for 48 h and 5 μmol/L As_2S_2 for 48 h,respectively, and (40.6±2.0)% cells did in combination treatment ( P <0.05). For U937 cells,the percentages of apoptotic cells were (6.0±1.1)% at 1 μmol/L STI571 for 48 h, (4.5±1.2)% at 5 μmol/L As_2S_2 for 48h, and (7.3±1.0)% in combination treatment. As_2S_2 decreased the bcr-abl fusion protein expression and PTK activity of c-abl and bcr-abl, but not for bcr-abl expression. Conclusion Combination treatment with As_2S_2 and STI 571 induced more apoptosis of K562 cells. The reduction of PTK activity may be involved in the mechanisms.

关 键 词：AS2S2 STI571 诱导 K562细胞 细胞凋亡 白血病 

分 类 号：R733.7[医药卫生—肿瘤]