吡柔比星诱导人膀胱癌T24细胞凋亡及其作用机制

Pirarubicin-induced apoptosis of bladder cancer T24 cells and the mechanism

目的 探讨吡柔比星（THP）对人膀胱癌T24细胞凋亡的影响及其作用机制.方法 使用不同浓度（0、0.1、1.0、10.0mg/L）THP作用膀胱癌T24细胞,逆转录-聚合酶链反应（RT-PCR）和Western blot检测细胞中生存素（Survivin）和半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3 mRNA及蛋白的表达;细胞计数试剂盒（CCK-8）检测细胞的增殖能力;流式细胞术检测细胞的周期及凋亡.结果 与空白组比较,各THP作用组T24细胞中Survivin mRNA及蛋白水平表达降低,Caspase-3 mRNA及蛋白水平表达增高（P＜0.01）;THP可抑制T24细胞的增殖,抑制率分别为（13.74±1.25）％、（36.28±0.75）％、（47.13±1.62）％,并诱导其凋亡,凋亡率为（15.29±0.98）％、（19.56±1.45）％、（23.81±0.81）％,并呈现一定的浓度和时间依赖性（P＜0.05）.结论 THP可促进膀胱癌T24细胞的凋亡,其可能通过抑制细胞中Survivin的表达,增强Caspase-3的活性而发挥作用.

Objective To investigate the pirarubicin （THP）-induced apoptosis of hunan bladder cancer T24 cells and its mechanisms.Methods T24 cells were cultured in vitro,and incubated with different concentrations of THP （0,0.1,1.0,and 10.0 mg/L）.The expression of Survivin and Cysteinyl aspartate-specific protease-3 （Caspase-3） was detected by reverse transcription-polymerase chain reaction （RT-PCR）,and conformed by Western blotting.The cell proliferation was measured by growth curve.The cell cycle and apoptosis rate of T24 cells were tested by flow cytometry.Results As compared with the control group,the expression of Survivin mRNA and protein was decreased,and that of Caspase-3 mRNA and protein increased in the THP-treated group （P 〈 0.01）.The inhibition rate was （13.74 ± 1.25） %,（36.28 ±0.75）% and （47.13±1.62）%,and the apoptosis rate was （15.29 ±0.98）%,（19.56± 1.45） % and （23.81 ± 0.81） in THP 0.1,1.0 and 10.0 mg/L groups respectively.THP inhibited the proliferation of T24 cells in a time-and concentration-dependent manner （P 〈 0.05）.Conclusion THP could inhibit the growth and induce apoptosis of T24 cells,which may be principally associated with the down-regulation of Survivin and up-regulation of Caspase-3.