小檗胺诱导K562细胞凋亡及其与bcr/abl gene及P210表达的关系

Apoptosis induced by berbamine in K562 cells correlates with the expression levels of bcr/abl gene and P210

目的 :探讨小檗胺 (BER)诱导K5 6 2细胞凋亡及其可能的机制。方法 :以流式细胞仪 (FCM)分析、电镜观察细胞微结构变化及基因组DNA电泳等方法检测细胞凋亡。以RT -PCR及Westernblot方法检测K5 6 2细胞BCR/ABLmRNA和蛋白质水平表达。结果 :FCM检测见到典型的凋亡峰 ,8 0mg/LBER作用 2 4 - 72h ,随药物作用时间延长 ,细胞凋亡率由 (2 9 2 0± 3 82 ) %上升至 (6 1 77± 4 35 ) % (P <0 0 1) ;以 2 0 - 8 0mg/LBER作用 2 4h ,随药物浓度增加 ,K5 6 2细胞的凋亡率也增加。电镜下可见明确的细胞凋亡的形态学改变。DNA电泳呈现典型的梯形条带。 16 0mg/LBER处理 2 4h ,K5 6 2细胞bcr/ablmRNA相对表达量及BCR/ABL融合蛋白质P2 10水平均明显低于对照组 ,分别为 0 73± 0 0 2vs 1 19± 0 0 2 (P <0 0 1)和 0 6 3± 0 0 1vs 1 0 4± 0 0 2 (P <0 0 1) ,呈时间 -浓度依赖效应。经BER作用后 ,K5 6 2细胞bcr/ablmRNA表达强度与P2 10水平呈正相关 (r =0 92 8,P <0 0 5 ) ,且细胞凋亡率与bcr/ablmRNA表达呈负相关 (r=- 0 997,P <0 0 1)。结论 :在体外BER对K5 6 2细胞有明显的促凋亡作用 。

AIM: To explore the effect of berbamine(BER) on apoptosis in K562 cells and its possible molecular mechanisms. METHODS: The apoptosis rate was measured by flow cytometry while electron microscopy and DNA electrophoresis were used to evaluate the characteristic changes of apoptosis, RT-PCR and Western blot were used to examine the expression levels of apoptosis related gene bcr/abl and BCR/ABL protein. RESULTS: By FCM, the apoptosis rate of K562 cells treated with 8.0 mg/L BER for 24 h and 72 h increased from (29.20±3.82)% to (61.77±4.35)% (P<0.01); The typical apoptosis morphologic changes and the DNA ladder were more clearly observed. After treated with BER 0 to 16.0 mg/L for 24 h, the expression levels of bcr/abl mRNA and P210 (semiquantity value) decreased quickly from 1.19±0.02 to 0.73±0.02 (P<0.01) and from 1.04±0.02 to 0.63±0.01 (P<0.01), respectively. CONCLUSION: BER induces apoptosis of K562 cells in a time-and concentration-dependent manner, the decline of bcr/abl mRNA and P210 may play an important role in the apoptotic effect of BER in K562 cells. BER could be used as a new clinical trials for bcr-abl^+ diseases such as CML.