冬凌草甲素对急性白血病HL-60细胞的增殖抑制作用及其作用机制  

Antiproliferation Effects of Oridonin on Leukemic HL-60 Cells and Its Mechanism

作　　者：刘加军[1] 林东军[1] 陆敏强[2] 张俊峰[2] 陈规划[2] 

机构地区：[1]中山大学附属第三医院血液内科,广州510630 [2]中山大学附属第三医院肝移植科,广州510630

出　　处：《热带医学杂志》2008年第10期1016-1019,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30772782);教育部新世纪优秀人才支持计划资助项目(No.NCET-06-0721);广东省中医药局课题(No.2007103)

摘　　要：目的探讨冬凌草甲素对急性白血病HL-60细胞的增殖抑制作用及其作用机制。方法以不同浓度的冬凌草甲素作用于体外培养的HL-60细胞,MTT法检测细胞生长抑制率,应用流式细胞仪检测细胞凋亡情况,瑞氏染色法观察细胞凋亡时的形态学变化。应用PCR-ELISA及RT-PCR法检测细胞凋亡前后端粒酶活性及端粒酶hTERTmRNA表达水平的变化。结果冬凌草甲素可显著的抑制HL-60细胞的生长,诱导细胞发生凋亡,并呈现出明显的量-效与时-效关系。冬凌草甲素作用48～60h后在瑞氏染色图片上可见核浓缩及核碎裂等典型的细胞凋亡特征,同时端粒酶hTERTmRNA的表达水平及端粒酶活性均明显降低。结论冬凌草甲素能抑制HL-60细胞的生长及诱导细胞发生凋亡;降低端粒酶hTERTmRNA的表达水平及端粒酶活性可能是其重要作用机制之一。Objective To investigate the antiproliferation effect of oridonin on leukemic HL-60 cells and it mechanism. Method HL-60 cells in culture medium were treated with different concentrations of oridonin. The inhibitory rate of the cells was measured by MTT assay. Cell apoptotic rate was detected by flow cytometry （FCM）. Morphology of cell apoptosis was observed by Wright＇s stain. Reverse transcriptase polymerase chain reaction （RTPCR） and PCR-enzyme-linked immunosorbent assay （ELISA） were used to detect hTERT mRNA expression and telomerase activity before and after apoptosis. Result Oridonin could inhibit the growth of HL-60 cells and cause apoptosis significantly. The suppression was in both time-dependent and dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by Wright＇s stain especially after the cells were treated 48 -60 h by oridonin. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently by treatment with oridonin in HL-60 cells. Conclusion Oridonin has apparent antiproliferation and apoptosis-inducing effects on HL-60 cells in vitro, downregulation of the hTERT mRNA expression and decreasing the telomerase activity of HL-60 cells may be its important antileukemia mechanisms.

关 键 词：冬凌草甲素 白血病 端粒酶 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤]