中药淫羊藿苷逆转肝癌HepG2.2.15细胞恶性表型及诱导分化研究  

Chinese medicine Icraiin reverses expression of hepatocarcinoma cell line HepG2.2.15 and induces differentiation

作　　者：王谦[1] 张玲[1] 毛海婷[1] 顾洪涛[1] 温培娥[1] 李翠玲[1] 杨尚军[2] 夏武青[3] 

机构地区：[1]山东省医学科学院基础医学研究所 山东省医药卫生肿瘤免疫与中药免疫重点实验室 山东省现代医用药物与技术重点实验室,山东省济南市250062 [2]山东省医学科学院药物研究所,山东省济南市250062 [3]山东省千佛山医院,山东省济南市250014

出　　处：《世界华人消化杂志》2007年第19期2087-2092,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金项目资助;No.30540041

摘　　要：目的:探讨淫羊藿苷(ICA)诱导HepG2.2.15细胞分化凋亡的作用机制.方法:MTT法检测细胞增殖;流式细胞术(FACS)检测细胞周期分布;RT-PCR方法检测P27基因、FLIP基因mRNA表达水平的变化;AnnexinⅤ-FITC/PI双染法检测ICA处理后HepG2.2.15细胞凋亡的变化;电化学发光法和速率散射比浊法分别检测ICA处理后HepG2.2.15细胞上清液中甲胎蛋白(AFP)和转铁蛋白(Tf)水平变化.结果:ICA作用HepG2.2.15细胞增殖呈抑制作用,具有时间依赖性.ICA处理后,HepG2.2.15细胞周期各时相分布与对照组相比发生变化,G0/G1期升高,S期减小,与对照组相比有显著性差异(56.26±1.56%vs49.68±1.34%,19.95±1.24%vs28.02±1.03%;P<0.01).ICA分别上调HepG2.2.15细胞P27和下调FLIP基因mRNA的表达水平(0.78vs0.27,0.54vs0.90);HepG2.2.15细胞凋亡率增加,AFP下降,Tf水平升高,与对照组相比均有显著性意义(7.09%vs0.59%,156±46mg/Lvs285±58mg/L,152.1±26mg/Lvs67.1±24mg/L;P<0.05).结论:ICA可能通过升高G0/G1期,减少S期抑制HepG2.2.15细胞的增殖;上调P27mRNA和Tf水平,下调FLIPmRNA的表达和AFP合成的水平来诱导细胞分化.AIM： To investigate the mechanism of Icraiin （ICA） induction of tumor cell differentiation and apoptosis in HepG2.2.15 cells.METHODS： MTT assay was used to explore the effects of ICA on the proliferation of HepG2.2.15 cells. Reverse transcriptase-polymerase chain reaction was used to examine mRNA levels of P27 and FLIP genes. Flow cytometry was used to detect apoptosis of HepG2.2.15 cells treated by Annexin V-FITC/PI. ABN ProSpec System nephelometer was used to examine alpha fetal protein （AFP）, and the Elecsys System was used for immunoassay detection of transferrin （Tf）.RESULTS： Treatment with ICA reduced the rate of proliferation in the HepG2.2.15 cell line. The inhibition of cell proliferation was time dependent （P 〈 0.05）. The distribution ratios of ICA-treated HepG2.2.15 cells were significantly different compared with those of the controls. The number of G0/Gl-phase cells increased （56.26 ± 1.56% vs 49.68 ± 1.34%, P 〈 0.01）, and S-phase cells decreased （19.95 ± 1.24% vs 28.02 ± 1.03%, P 〈 0.01）. ICA increased the relative expression level of P27 mRNA from 0.27 to 0.78, and re- duced that of FLIP mRNA from 0.90 to 0.54. ICA also decreased AFP expression level from 285 ± 58 to 156 ± 46 rag/L, while it increased that of Tf from 67.1± 24 to 152.1 ± 26 mg/L （P 〈 0.05）.CONCLUSION： ICA significantly inhibited HepG2.2.15 cell proliferation by affecting the cell cycle distribution ratios, and by reducing the number of S-phase cells and increasing G0/G1- phase cells. ICA could enhance the expression levels of P27 mRNA and Tf, while simultaneously inhibiting the tumor marker AFP and in- ducing HepG2.2.15 cell differentiation.

关 键 词：淫羊藿苷 HEPG2.2.15 P27 甲胎蛋白 转铁蛋白 FLIP 

分 类 号：R735.7[医药卫生—肿瘤]