硼替佐米对K562细胞增殖、凋亡及SHIP基因表达的影响

本研究旨在探讨蛋白酶体抑制剂硼替佐米(bortezomib)对K562细胞的增殖、凋亡及SHIP基因表达的影响。将不同浓度的硼替佐米作用于K562细胞,采用MTT法测定细胞增殖活性,用流式细胞术检测细胞凋亡,RT-PCR方法检测SHIP mRNA表达。结果表明,硼替佐米10、20、50和100 nmol/L作用于K562细胞24 h,细胞增殖抑制率分别为(5.76±1.47)%、(10.55±1.59)%、(17.14±2.05)%和(27.69±3.57)%,空白对照组为(1.30±0.10)%;20nmol/L硼替佐米作用于K562细胞24、48、72 h,细胞增殖抑制率分别为(10.55±1.59)%、(16.33±2.53)%、(19.78±1.56)%,24 h组与48 h组比较,差异具有统计学意义(P<0.05),10、20、50、100 nmol/L硼替佐米作用于K562细胞24h,Annexin V-FITC/PI双标法显示其凋亡率分别为(12.7±0.6)%、(26.9±0.9)%、(32.6±1.2)%、(72.5±1.5)%,均高于对照组(1.0±0.5)%(P<0.05)。RT-PCR法检测结果显示应用硼替佐米作用后SHIP mRNA表达上调明显,与空白对照组比较差异有统计学意义。结论:硼替佐米呈时间、浓度依赖性抑制K562细胞增殖,并能通过上调SHIP基因的表达诱导细胞凋亡。

Effect of Proteasome Inhibitor Bortezomib on Proliferation, Apoptosis and SHIP gene expression in K562 cells

This study was aimed to investigate the effects of proteasome inhibitor bortezomib on proliferation,apoptosis and the SHIP expression of K562 cells.K562 cells were treated with bortezomib of different concentrations.Cell proliferation was analyzed by MTT assay,cell apoptosis was detected by flow cytometry and SHIP mRNA expression was assayed by RT-PCR.The results showed that after being treated with 10,20,50 and 100 nmol/L bortezomib for 24 h,the inhibitory rates of K562 cells were（5.76±1.47）%,（10.55±1.59）%,（17.14±2.05）% and（27.69±3.57）% respectively,and were higher than that in control（1.30±0.10）;when K562 cells were treated with 20 nmol/L bortezomib for 24,48 and 72 h,the inhibitory rates of cell proliferation were（10.55±1.59）%,（16.33±2.53）% and（19.78±1.56）% respectively,there was statistic difference of cell proliferation rate between 24 h group and 48 h group（P0.05）.After being treated with 10,20,50,100 nmol/L bortezomib for 24 h,the apoptotic rates of K562 cells were（12.7±0.6）%,（26.9±0.9）%,（32.6±1.2）% and（72.5±1.5）% respectively,and all hyher than that in contol（1.0±0.5）%（P0.05）.According to results of RT-PCR detection,the expression level of SHIP mRNA was obviously up-regulated after treatment with bortezomin,and showed statistical difference in comparison with control.It is concluded that bortezomib inhibits proliferation of K562 cells in time and concentration-tependent manner and induces apoptosis through up-regulation of SHIP gene.