D-柠檬烯诱导人膀胱癌细胞周期阻滞及凋亡的研究

目的探讨D-柠檬烯对人膀胱癌EJ细胞增殖、细胞周期分布以及凋亡的影响。方法用XTT法、流式细胞技术(碘化丙啶)和流式细胞仪AnnexinV法对D-柠檬烯处理后EJ细胞增殖、周期分布以及凋亡率进行检测。结果经2、4mmol/L浓度D-柠檬烯处理后,EJ细胞增殖活性(OD值)分别为0.801±0.016和0.148±0.008,与对照组(1.218±0.031)比较,生长受到明显抑制,差异有统计学意义(P0.05)。细胞周期分析显示,给药组的合成期(S)细胞百分比为23.05%,而未给药的对照组为38.59%,两组差异有统计学意义(P0.05)。另外,D-柠檬烯作用24h后的EJ细胞出现典型的亚二倍体凋亡峰,药物浓度越高凋亡细胞越多,与对照组比较,差异有统计学意义(P0.05)。结论 D-柠檬烯对人膀胱癌EJ细胞具有显著的抑制作用;它还能使EJ细胞停滞于S期,并能诱导细胞凋亡。

D-Limonene induced cell cycle perturbations and apoptosis in human bladder cancer lines

【Objective】To investigate the modulating effects of D-Limonene on the cell cycle and apoptosis of human bladder cancer lines.【Methods】The cytotoxic activity and apoptosis were analyzed by XTT and Annexin V assay as well as DNA cytometry,respectively.The effects on the cell cycle were assessed by flow cytometry of propidium iodide.【Results】The proliferation of EJ cell was inhibited by exposure to 2mmol/L and 4mmol/L of D-limonene for 48 hours,and the optical density（OD） was 0.801 ± 0.016 and 0.148 ± 0.008,respectively.There were statistical differences between the experimental groups and the control group（1.218 ± 0.031）（P〈 0.05）.D-Limonene inhibited cell proliferation in dose dependent manner.Cell cycle analysis showed 23.05% of cell percentage of synthesis period in medication administration team,while 38.59% in the control group,significant difference was available（P〈 0.05）.The classic hypodiploid apoptotic peak was occurred in EJ cell diposed with D-limonene 24h later,and the amount of apoptosis cell rised with the drug concentration.The significant difference was available contrast with the control group（P〈 0.05）.【Conclusion】D-limonene obviously suppresses the growth of EJ cell lines.The cytoxicity depends on the dosage of d-limonene.Moreover,d-limonene can stagnate EJ cell lines in S period,and introduce the cell apoptosis.Our study demonstrated that d-limonene exerts its cytotoxic effect on EJ cell lines by inducing cell cycle perturbations and apoptosis.