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目的:探讨硼替佐米诱导慢性粒细胞白血病细胞株K562细胞凋亡及新基因bcl2l12在其中的作用。方法:MTT比色法观察硼替佐米对K562细胞的生长抑制作用;Annexin-V标记和线粒体跨膜电位(Δψm)分析细胞凋亡;RT-PCR方法检测0、6、12和24hfas、bcl2l12、bcl-2、bim、bax、caspase-3和caspase-9基因表达变化。结果:硼替佐米抑制K562细胞生长呈时间和剂量依赖性,24h和48h半数抑制浓度分别为161.41nmol/L和96.33nmol/L;硼替佐米诱导K562细胞凋亡,12h Annexin-V阳性细胞就开始增高,并呈时间依赖性,Δψm减低;RT-PCR显示fas、bcl2l12、caspase-3和caspase-9表达增高,但bcl-2、bim和bax表达无明显改变。结论:硼替佐米可以抑制K562生长并诱导凋亡,上调fas、bcl2l12,使线粒体膜电位下降,激活caspase-9和caspase-3基因,促使DNA发生断裂可能是其诱导凋亡的机制之一。
OBJECTIVE: To investigate the apoptosis of B5 cells induced by bortezomib and the role of bcl2l12 in the treatment of chronic myelogenous leukemia cell line K562. Methods MTT assay was used to observe the inhibitory effect of bortezomib on the growth of K562 cells. Annexin-V and mitochondrial transmembrane potential (Δψm) were used to detect apoptosis. RT-PCR was used to detect the expression of bcl21, bcl-2, bim, bax, caspase-3 and caspase-9 gene expression changes. RESULTS: Bortezomib inhibited the growth of K562 cells in a dose- and time-dependent manner. The half-inhibitory concentrations were 161.41 nmol / L and 96.33 nmol / L at 24h and 48h, respectively. Bortezomib induced the apoptosis of K562 cells. Annexin-V positive cells The expression of fas, bcl2l12, caspase-3 and caspase-9 increased, but the expression of bcl-2, bim and bax did not change significantly. CONCLUSION: Bortezomib can inhibit the growth of K562 cells and induce apoptosis. The up-regulation of fas, bcl2l12, the decrease of mitochondrial membrane potential and the activation of caspase-9 and caspase-3 genes may promote the apoptosis of K562 cells.