Z-ajoene引起HL-60细胞G_2/M期阻滞及端粒酶表达活性的降低  

Z-ajoene causes cell cycle arrest at G_2/M and decrease of telomerase activity in HL-60 cells

作　　者：叶颖[1] 杨华瑜[1] 吴军[1] 李敏[1] 闽吉梅[1] 崔景荣[1] 

机构地区：[1]北京大学医学部天然药物及仿生药物国家重点实验室,100083

出　　处：《中华肿瘤杂志》2005年第9期516-520,共5页Chinese Journal of Oncology

摘　　要：目的探讨Z-ajoene诱导细胞周期阻断于G2/M期的分子机制以及对端粒酶活性的抑制作用。方法采用MTT法测定Z-ajoene对HL-60细胞的增殖抑制率;以流式细胞仪测定Z-ajoene对HL-60细胞的周期阻断作用,并采用Westernblot法测定相关蛋白p34cdc2、cyclinB1、cyclinA的表达水平;TRAP-银染法测定端粒酶活性的变化,同时采用RT-PCR方法检测端粒酶hTRT和TP1mRNA的表达。结果10μmol/LZ-ajoene作用后能明显将HL-60细胞阻滞于G2/M期。在药物作用10h后,G2/M期细胞的比例达到顶峰,与对照组比较增加了1.95倍。另外,10μmol/LZ-ajoene作用后出现cyclinB1蛋白的聚集及p34cdc2蛋白表达的降低,但cyclinA蛋白的水平并无显著性变化。经Z-ajoene作用24h后,HL-60细胞端粒酶活性显著降低。而不同浓度的药物作用12h,其端粒酶活性均无显著性变化。在10μmol/LZ-ajoene作用24h后,能够降低端粒酶hTRT和TP1mRNA的水平。结论Z-ajoene能够在作用较短时间内明显将细胞周期阻断于G2/M期,同时伴有cyclinB1聚集及p34cdc2的抑制;而HL-60细胞的端粒酶活性仅在药物作用较长时间后才有一定降低,并且端粒酶hTRT和TP1mRNA的表达也同时降低。结果表明,Z-ajoene的细胞周期阻断作用可能是造成端粒酶缺失及HL-60细胞生长抑制的重要原因。Objective To investigate the molecular mechanisms of Z-ajoene mitosis blocking and telomerase inhibitory effects on HL-60 cells. Methods Proliferation inhibition of HL-60 cell line was evaluated by MTF assay. Z-ajoene-induced mitotic blocking effect was investigated by flow cytometry. Immunoblotting analysis was used to determine cell cycle regulatory proteins. The telomerase activity of HL- 60 cells was detected by TRAP-silver stain assay. Telomerase hTRT and TP1 mRNA level were determined by RT-PCR. Results Z-ajoene displayed great proliferation inhibiting effect on HL-60 cells. Progressive increase in the percentage of mitotic block at G2/M phase was observed from 4 h to 12 h after treatment with 10 μmol/L Z-ajoene, with a peak at 10 h, which was 1.95 times higher than that in control. Z-ajoene also caused an increase in cyclin B1 accumulation and a decrease of p34^cdc2 expression. But Z-ajoene did not change the level of cyclin A. After treating with 10 μmol /L Z-ajoene for 24 h, the telomerase activity of HL-60 cells was also decreased in a dose-independent manner. Furthermore, telomerase hTRT and TP1 mRNA levels decreased after 10 μmol/L Z-ajoene treatment for 24 h. Conclusion The results suggest that Z-ajoene has potent anti-cancer activity, and that its inhibitory effect on telomerase activity and on cell growth might be the result of G2/M phase blocking.

关 键 词：Z-ajoene HL-60细胞 端粒酶 细胞周期 G2/M期阻滞 细胞端粒酶活性 cyclinB1蛋白 表达活性 P34^CDC2 端粒酶HTRT 

分 类 号：R733.7[医药卫生—肿瘤]