姜黄素抑制人肝癌细胞BEL-7402中VEGF和HIF-1α表达通过PI3K／AKT／FRAP信号传导通路  

Curcumin inhibit the expression of VEGF and HIF-1α protein in human hepatocellular carcinoma BEL-7402 cells by PI3K/AKT/ FRAP signal transduction pathway

作　　者：孙军[1] 李岩[2] 王贺玲[2] 

机构地区：[1]辽宁省锦州市中心医院消化内科,辽宁锦州121004 [2]中国医科大学盛京医院消化内科

出　　处：《中国医师杂志》2008年第1期41-44,共4页Journal of Chinese Physician

摘　　要：目的探讨MAPK和PI3K信号传导通路在姜黄素调节VEGF和HIF-1α表达中的作用。方法分别加入LY29400225μmol／L、50μmol／L，U012610μmol／L、20μmol／L，rapamycin 5ng／ml、10ng／ml处理人肝癌细胞BEL-7402，30min后加入姜黄素10μmol／L，对照组单独加入0、10μmol／L姜黄素，缺氧环境中培养6h后，行RT—PCR和Western Blot检测VEGF蛋白、mRNA和HIF-1α蛋白表达；分别加入不同浓度的的姜黄素以及LY294002 25 μmol／L处理人肝癌细胞BEL-7402，缺氧环境中培养6h后，行Western Blot检测磷酸化AKT和总AKT蛋白表达。结果姜黄素10μmol／L＋LY29400225μmol／L组、姜黄素10μmol／L＋LY294002 50 μmol／L组、姜黄素10μmol／L＋rapamycin 5 ng／ml组、姜黄素10μmol／L＋rapamycin 10 ng／na组HIF-1α蛋白、VEGF蛋白、mRNA表达分别与姜黄素10μmol／L组相比降低，差异有统计学意义（P〈0．01）；而姜黄素10μmol／L＋U012610μmol／L组、姜黄素10μmol／L＋U0126 20 μmol／L组HIF—1α蛋白、VEGF蛋白、mRNA表达分别与姜黄素10μmol／L组相比差异无统计学意义（P〉0.05）；不同浓度的姜黄素、LY294002 25 μmol／L处理的人肝癌细胞BEL-7402缺氧6h后磷酸化AKT蛋白表达逐渐降低，LY294002 25 μmol／L可以基本阻断磷酸化AKT蛋白的表达，而对总AKT蛋白表达无明显变化。结论姜黄素对人肝癌细胞BEL-7402中HIF-1α蛋白和VEGF的表达通过P13K／AKT／FRAP信号传导通路。Objective To investigate the effects of PI3K and MAPK signal transduction pathway on the expression of VEGF and HIF-1α in human hepatocellular carcinoma BEL-7402 cells. Methods Human hepatocellular carcinoma BEL-7402 cells were pretreated with 20μmol/L, 50μmol/L of LY294002, 10μmol/L, 20μmol/L of UO126,5ng/ml, 10ng/ml of rapamycin for 30 minutes, followed by treatment with 10μmol/L of cureumin and alone treated with 0 or 10μmol/L of cureumin in control group, the cells were cultured under hypoxic conditions for 6 hours. The expression of VEGF, mRNA and HIF-1α were detected by RT-PCR and Western Blot. Cells were treated with different concentrations of cureumin and 25μmol/L of LY294002 under hypoxic conditions for 6 hours, the phosphorylation of AKT and total AKT were detected by Western Blot. Results The expression of VEGF protein, mRNA and HIF-1α protein were significantly different among 10μmol/L of curcumin plus 25 μmol/L of LY294002 group, 10μmol/L of curcumin plus 50μmol/L of LY294002 group, 10μmol/L of cureumin plus 5ng/ml rapamycin group and 10μmol/L of curcumin plus 10ng/ml rapamycin group, compared with 10μmol/L of curcumin group （ P 〈 0. 01 ）. The expression of VEGF protein, mRNA and HIF-1 α protein were no significantly different among 10μmol/L of cureumin plus U0126 10μmol/L group, 10μmol/L of cureumin plus U0126 20μmol/L group and 10μmol/L of curcumin group （ P 〉 0. 05）. The expression of phosphorylation of AKT protein in human hepatocellular carcinoma BEL-7402 cells treated with different concentrations of curcumin and 25μmol/L of LY294002 under hypoxic conditions for 6 hours decreased gradually. 25μmol/L of LY294002 can block the expression of the phosphorylated AKT protein, but it can not influence on the expression of the total AKT protein. Conclusions Cureumin inhibit the expression of VEGF and HIF-1α protein in human hepatocellular carcinoma BEL-7402 cells by PI3K/AKT/FRAP signal transduction pathway.

关 键 词：姜黄素/药理学 肝肿瘤 血管内皮生长因子类 缺氧 丝裂原激活蛋白激酶激酶类 1-磷脂酰肌醇3-激酶 

分 类 号：R735.7[医药卫生—肿瘤]