丹参酮ⅡA对肝癌SMMC-7721细胞COX-2表达的影响  

Effect of tanshinone ⅡA on COX-2 expression in hepatocellular carcinoma cell line SMMC-7721

作　　者：徐俊丽[1] 和水祥[1] 陈静宏[2] 符寒[1] 赵刚[1] 王艳丽[1] 任牡丹[1] 

机构地区：[1]西安交通大学医学院第一附属医院消化内科,陕西省西安市710061 [2]西安交通大学医学院病原微生物与寄生虫系,陕西省西安市710061

出　　处：《世界华人消化杂志》2006年第14期1352-1356,共5页World Chinese Journal of Digestology

基　　金：陕西省中医局科研基金项目;No.2001-040~~

摘　　要：目的:观察丹参酮ⅡA对肝癌SMMC-7721细胞生长和凋亡的影响及其作用机制．方法:体外培养肝癌SMMC-7721细胞株,经丹参酮ⅡA(终浓度0．5 mg／L)作用后,采用四唑盐(MTT)比色法检测细胞增殖,透射电镜观察细胞凋亡,流式细胞仪检测细胞凋亡,免疫细胞化学SABC法检测COX-2蛋白表达,放射免疫法检测前列腺素E2(PGE2)含量．结果:丹参酮ⅡA对肝癌细胞的生长有明显的抑制作用,并呈剂量依赖性．以0．5 mg／L作用终浓度抑制作用最明显,其48 h的抑制率为 69．3％,与对照组相比差异有显著性(P<0．01)．电镜下观察,丹参酮ⅡA作用后肝癌细胞表现为细胞皱缩、核染色质浓缩、核碎裂以及凋亡小体形成等凋亡特征性的形态改变．5 mg/ L丹参酮ⅡA作用后,随时间的延长,凋亡率逐渐升高,48 h达到高峰,随后逐渐下降(24,48, 72 h凋亡率分别为7．45％±0．33％、6．59％± 0．45％、4．78％±1．05％),与对照组比较,各处理组都有显著性差异(均P<0．01),丹参酮作用组肝癌细胞COX-2表达明显减少,其培养液中 PGE2的产生量下降,与对照组相比差异均有显著性(P<0．01)．结论:丹参酮ⅡA可能是通过下调COX-2 mRNA的表达水平发挥其对肝癌细胞生长抑制及促进凋亡作用．AIM： To observe the effect of Tanshinone Ⅱ A on the proliferation and apoptosis of human hepatocellular carcinoma cell line SMMC-7721 and its machanism. METHODS： The human hepatocellular carcinoma cell line SMMC-7721 cultured in vitro was treated with different concentrations （final 0.5 rag/L） Tanshinone Ⅱ A. The proliferation of the cells was measured by MTT assay, and the apoptosis of the cells was investigated by flow cytometry and transmission electron microscopy （TEM）. The expression of Cyclooxygenase-2 （COX-2） was detected by immunocytochemistry. The levels of prostaglandin E2 （PGE2） in medium were measured by radioimmunoassay RESULTS： Tanshinone Ⅱ A inhibited the growth of SMMC-7721 cells in a doseependent manner. The inhibitory rate reached the peak （69.3%） after 0.5 mg/L Tanshinone Ⅱ A was used for 48 h, which was significantly higher than that in the controls （P 〈 0.01）. Typical features of apoptosis as cell shrinkage, nuclear chromatin condensation, fragmentation, and the formation of apoptotic bodies was found by TEM in the cells treated with Tanshinone II A. FCM analysis showed the apoptotic rates for 24, 48 and 72 h were 7.45% ± 0.33%, 6.59% ± 0.45%, and 4.78% ± 1.05%, respectively, when 5 mg/L Tanshinone Ⅱ A was used, which were markedly higher than those in the controls （all P 〈 0.01）. Both the expression of COX-2 （P 〈 0.01） and the production of PGE2 （40.18 ± 1.37μg/L vs 75.31 ±1.64μg/L, P 〈 0.01） were decreased significantly after TanshinoneⅡ A treatment as compared with those in the controls. CONCLUSION： Tanshinone Ⅱ A inhibits the proliferation and increases the apoptosis of human hepatocellular carcinoma cell line SMMC-7721 by down-regulation of COX-2 protein expression.

关 键 词：丹参酮 环氧合酶-2 肝癌细胞 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]