柔红霉素对人B细胞淋巴瘤OCI-LY7细胞凋亡的诱导作用及其机制

Inductive effect of daunorubicin on apoptosis of human B-cell lymphoma OCI-LY7 cells and its mechanism

目的:探讨柔红霉素对人B细胞淋巴瘤OCI-LY7细胞凋亡的影响及其机制,阐明柔红霉素在弥漫大B细胞淋巴瘤中的耐药机制。方法:将体外培养的OCI-LY7细胞分为0、0.1、1.0和10.0 mg·L^-1柔红霉素组。采用MTT法检测各组OCI-LY7细胞存活率,流式细胞术检测各组OCI-LY7细胞凋亡率,Western blotting法检测各组OCI-LY7细胞中凋亡相关蛋白Bax、cleaved caspase-3和survivin表达水平。后续实验分为对照组、柔红霉素组、柔红霉素+空载体组和柔红霉素+survivin组,按照上述方法检测survivin过表达后各组OCI-LY7细胞存活率、凋亡率和细胞中凋亡相关蛋白表达水平。结果:与0 mg·L^-1组比较,处理48 h后0.1、1.0和10.0 mg·L^-1柔红霉素组OCI-LY7细胞存活率和细胞中survivin蛋白表达水平逐渐降低(P<0.05),而细胞凋亡率和细胞中cleaved caspase-3及Bax蛋白表达水平逐渐升高(P<0.05)。与对照组比较,柔红霉素组OCI-LY7细胞存活率和细胞中survivin蛋白表达水平明显降低(P<0.05),而细胞凋亡率和细胞中cleaved caspase-3及Bax蛋白表达水平明显升高(P<0.05);与柔红霉素组比较,柔红霉素+survivin组OCI-LY7细胞存活率和细胞中survivin蛋白表达水平明显升高(P<0.05),而细胞凋亡率和细胞中cleaved caspase-3及Bax蛋白表达水平明显降低(P<0.05)。结论:柔红霉素可诱导OCI-LY7细胞凋亡,其作用机制可能与下调survivin蛋白表达有关。

Objective:To investigate the effect of daunorubicin on the apoptosis of human B-cell lymphoma OCI-LY7 cells and its mechanism,and to elucidate the drug resistance mechanism of daunorubicin in the diffuse large B-cell lymphoma.Methods:The OCI-LY7 cells cultured in vitro were divided into 0,0.1,1.0 and 10.0 mg·L^-1 daunorubicin groups.The survival rates of OCI-LY7 cells in various groups were detected by MTT assay,the apoptotic rates of the OCI-LY7 cells in various groups were detected by flow cytometry,and the expression levels of apoptosis-relatedProteins Bax,cleaved caspase-3,and survivin in the OCI-LY7 cells in various groups were detected by Western blotting method.The follow-up experiments were divided into control group,daunorubicin group,daunorubicin+vector group and daunorubicin+survivin group;the survival rates,the apoptotic rates and the expression levels of apoptosis-relatedProteins in the OCI-LY7 cells in various groups after overexpression of survivin were detected by the above-mentioned methods.Results:Compared with 0 mg·L^-1 group,the survival rates of OCI-LY7 cells and the expression levels of survivinProtein in the OCI-LY7 cells in 0.1,1.0 and 10.0 mg·L^-1 daunorubicin groups were decreased gradually after treated for 48 h(P<0.05),while the apoptotic rates and the expression levels of Cleaved caspase-3 and BaxProteins in the OCI-LY7 cells were gradually increased(P<0.05).Compared with control group,the survival rate and the expression level of survivinProtein in the OCI-LY7 cells in daunorubicin group were significantly decreased(P<0.05),while the apoptotic rate and the expression levels of cleaved caspase-3 and BaxProteins in the OCI-LY7 cells were significantly increased(P<0.05);compared with daunorubicin group,the survival rate and the expression level of survivinProtein in the OCI-LY7 cells in daunorubicin+survivin group were significantly increased(P<0.05),while the apoptotic rate and the expression levels of cleaved caspase-3 and BaxProtein in the OCI-LY7 cells in daunorubicin+vector group were significantly decreased(P<0.05).Conclusion:Daunorubicin can induce the apoptosis of OCI-LY7 cells,and its mechanism may be related to down-regulation of the expression of survivinProtein.