摘要
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。
AIM： To detect the treatment of K562 leukemia cells with bortezomib altering the expression of genesfas, bcl-2, bcl2112, bim, bax, caspase -9 and caspase -3. METHODS： MTT assay was used to detect the inhibition of proliferation. Apoptosis was detected by Annexin - V staining and mitochondrial transmembrane potential （△ψm）. RT - PCR was used to analyze the mRNA expressions offas, bcl - 2, bcl2112, bim, bax, caspase - 3 and caspase - 9. RESULTS： Bortezomib caused a time - and dose - dependent inhibition of cell proliferation and IC50 of 24 h and 48 h were 161.41 nmol/L and 96. 33 nmol/L, respectively. At the concentration of 104 nmol/L, bortezomib induced apoptosis in a timedependent manner, including increasing annexin -V poSitivity and decreasing the △ψm. RT- PCR showed that bortezomib up- regulated the mRNA expression offas, bcl2112, caspase- 9 and caspase- 3, but mRNA expressions of bcl - 2, bim and bax did not changed obviously. CONCLUSION： Bortezomib inhibits the proliferation of K562 and induces apoptosis, in which fas, bcl2112, caspase - 9 or caspase - 3 gene is one of the main genes taking part in.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第5期883-887,共5页
Chinese Journal of Pathophysiology
基金
宁波市医学科技计划资助项目(No.200520)
