张俊峰;陈规划;陆敏强;李华;蔡常洁;杨扬;
ZHANG Jun-feng,CHEN Gui-hua,LU Ming-Qing,LI Hua,CAI Chang-Jie,YANG Yang(Department of Liver Transplantation,The Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

• 1:中山大学附属第三医院肝移植中心
• 2:中山大学附属第三医院肝移植中心
• 3:中山大学附属第三医院肝移植中心
• 4:中山大学附属第三医院肝移植中心

摘要(Abstract)：

目的:探讨冬凌草甲素诱导人肝癌细胞BEL-7402凋亡中Bc l-2表达及端粒酶活性的变化。方法:以8,16,24,32μmol.L-1不同浓度的冬凌草甲素作用于体外培养的BEL-7402细胞,应用流式细胞仪检测细胞凋亡,Hoechst 33258荧光染色法。W estern b lot观察Bc l-2,Bax表达变化。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:冬凌草甲素可诱导肝癌细胞BEL-7402发生细胞凋亡,具有明显的量-效与时-效关系。冬凌草甲素作用60 h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。凋亡过程中Bc l-2蛋白表达的降低和Bax蛋白上调,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:冬凌草甲素在诱导肝癌细胞BEL-7402发生细胞凋亡过程中,能降低端粒酶hTERT mRNA的表达水平和端粒酶活性,同时抗凋亡蛋白Bc l-2表达的降低和促凋亡蛋白Bax上调。
Objective: To investigate the change of Bcl-2 expression and telomerase during the apoptosis induced by oridonin on human hepatocelluar carcinoma BEL7402 cells.Method:BEL-7402 cells in culture medium were given 8,16,24,32 μmol·L^(-1) different concentrations of oridonin.The cell apoptotic rate was detected by flow cytometry(FCM),morphology of cell apoptosis was observed by Hoechst 3325 staining.Bcl-2 and Bax expressions were detected by Western blotting.Reverse transcriptase polymerase chain reaction(RT-PCR) and PCR enzyme-linked immunosorbent assay(ELISA) were used to detect hTERT mRNA expression and telomerase activity.Result: Oridonin induced BEL-7402 cells apoptosis significantly,and the apoptosis rate was both in time-and dose-dependent manner.Marked morphological changes of cell apoptosis were observed very clearly by Hoechst 33258 staining after the cells exposed to oridonin for 60 hours;Western blotting showed that Bcl-2 expression was down-regulated and Bax expression up-regulated concurrently along with the apoptotic process,and the expression of hTERT mRNA as well as activity of telomerase were decreased concurrently.Conclusion: Oridonin could decrease the expression of hTERT mRNA and telomerase activity as well as down-regulation of Bcl-2 and up-regulation of Bax expression during the apoptosis of BEL-7402 cells.

关键词(KeyWords)： 冬凌草甲素;肝癌;细胞凋亡;Bcl-2;Bax;端粒酶
oridonin;hepatocellular carcinoma;apoptosis;Bcl-2;Bax;telomerase

Abstract:

Keywords:

基金项目(Foundation): 国家重点发展研究计划(973)资助项目(2003CB515507)

作者(Authors): 张俊峰;陈规划;陆敏强;李华;蔡常洁;杨扬;
ZHANG Jun-feng,CHEN Gui-hua,LU Ming-Qing,LI Hua,CAI Chang-Jie,YANG Yang(Department of Liver Transplantation,The Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

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