摘要
目的：探讨冬凌草甲素对肝癌BEL-7402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BEL-7402细胞，MTY法检测细胞生长抑制率，应用流式细胞仪检测细胞凋亡，Hoechst 33258荧光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hTERT mRNA表达水平的变化。结果：冬凌草甲素可显著的抑制BEL-7402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Hoechst 33258荧光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hTERT mRNA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BEL-7402细胞的生长及诱导细胞发生凋亡；降低端粒酶hTERT mRNA的表达水平及端粒酶活性可能是其重要作用机制之一。
AIM： To investigate the antiproliferation effects of oridonin on hepatocellular carcinoma BEL-7402 cells and its mechanisms of action. METHODS： BEL-7402 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 Hoechst 33258 stain. Reverse transcriptase polymerase chain reaction （RT-PCR） and PCR-enzyme-linked immunosorbent assay （ELISA） were used to detect hTERT mRNA expression and telomerase activity before and after apoptosis. RESULTS： Oridonin could inhibit the growth of BEL-7402 cells and cause apoptosis significantly. The suppression was in both time-dependent and dose-dependent manner. Marked morphological changes in cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by Hoechst 33258 stain especially after the cells were treated 48-60 h by oridonin. The expression of hTERT mRNA as well as activity of telomerase decreased concurrently by treatment with oridonin in BEL-7402 cells. CONCLUSION： Oridonin has apparent antiproliferation and apoptosis - inducing effects on BEL-7402 cells in vitro, downregulation of the hTERT mRNA expression and decreasing the te- lomerase activity of BEL-7402 cells may be one of its important anti-hepatocellular carcinoma mechanisms.
出处
《中成药》
CAS
CSCD
北大核心
2006年第9期1325-1329,共5页
Chinese Traditional Patent Medicine
基金
国家重点发展研究计划(973)资助项目(编号2003CB515507)
关键词
冬凌草甲素
肝癌
端粒酶
细胞凋亡
oridonin
hepatocellular carcinoma
telomerase
apoptosis
