水翁花黄酮DMC诱导肝癌细胞SMMC-7721凋亡的机制研究  

Study on mechanism of inducing apoptosis in human hepatoma SMMC-7721 cells by DMC,a chalcone from buds of Cleistocalyx operculatus

作　　者：叶春林[1] 赖依峰[1] 刘宣淦[1] 黄琦[1] 

机构地区：[1]浙江科技学院生物与化学工程学院,浙江杭州310023

出　　处：《中国中药杂志》2014年第15期2942-2946,共5页China Journal of Chinese Materia Medica

基　　金：浙江省自然科学基金项目(LY12C02002)

摘　　要：目的:研究水翁花黄酮类化合物2',4'-二羟基-6'-甲氧基-3',5'-二甲基查耳酮(DMC)体外诱导人肝癌细胞SMMC-7721凋亡作用及其分子机制。方法:采用改进的Sellins-Cohen方法检测不同浓度的DMC对SMMC-7721细胞DNA片段化的影响。用PCR-ELISA法测定肿瘤细胞的端粒酶活性,用半定量RT-PCR法测定c-myc和人端粒酶逆转录酶(hTERT)的mRNA表达量,Western blot法检测c-myc及hTERT蛋白的表达。结果:DMC作用于人肝癌SMMC-7721细胞后,引起SMMC-7721细胞DNA片段化率增大,片段化率效果有明显的浓度依赖性。与对照组相比,20μmol·L-1DMC处理48 h后,肝癌细胞的端粒酶活性下降了(66.2±2.1)%。c-myc和hTERT的mRNA表达水平分别下降了(67.3±2.1)%,(64.4±2.3)%,c-myc和hTERT的蛋白表达水平分别下降了(69.6±1.9)%,(71.3±2.4)%。结论:DMC能诱导SMMC-7721细胞凋亡,其凋亡机制可能与c-myc基因和hTERT的mRNA以及蛋白的表达降低有关。Objective：To study the in-vitro inducing apoptosis mechanism of human hepatoma SMMC-7721 cells by 2＇,4＇-dihydroxy-6＇-methoxy-3＇,5＇-dimethylchalcone（DMC）,a chalcone compound from Cleistocalyx operculatus.Method：Quantitative DNA fragmentation assay was carried out to detect the effect of DMC of different concentrations on SMMC-7721 cells,according to the method of Sellins and Cohen with some modifications.Telomerase activities of the cells were determined by PCR-ELISA methods.The expression quantity of c-myc and hTERT mRNA were determined by semi-quantitative RT-PCR.The effect of DMC on expression levels of cmyc and hTERT protein were measured by western blot.Result：The percentage of DNA fragmentation increased with notable concentration dependence,after treatment with DMC for 48 h.Compared with that of control group,the telomerase activity of the cells decreased by（66.2 ± 2.1） % after 48 h treatment with 20 μmol·L- 1 DMC,the mRNA expression of c-myc and hTERT decreased by（67.3 ±2.1） % and（64.4 ±2.3） %,respectively,and the protein expression of c-myc and hTERT decreased by（69.6 ±1.9） % and（71.3 ± 2.4） %,respectively.Conclusion：DMC can induce SMMC-7721 cell apoptosis and the apoptosis mechanism may be related to the decreased mRNA and protein expression of c-myc and hTERT.

关 键 词：水翁花 凋亡 黄酮 肿瘤 端粒酶 

分 类 号：R285[医药卫生—中药学]