黄荆子木脂素3对人肝癌HepG2细胞增殖的抑制作用及对蛋白激酶Akt、ERK1/2信号通路的影响  

Effects of Vitex negundo Lignans-3 on the Proliferation Inhibition of Human Hepatic Cancer HepG2 Cells and Protein Kinase Akt and ERK1/2 Signal Pathway

作　　者：李一春[1] 杨文军[2] 陈艳 

机构地区：[1]南华大学附属第二医院妇产科,湖南衡阳421001 [2]南华大学附属第二医院普外科,湖南衡阳421001

出　　处：《中国药房》2013年第33期3099-3101,共3页China Pharmacy

摘　　要：目的:研究黄荆子木脂素3对人肝癌HepG2细胞增殖的抑制作用,及其与丝氨酸-苏氨酸蛋白激酶(Akt)、细胞外调节蛋白激酶1/2(ERK1/2)信号通路的关系。方法:以人肝癌HepG2细胞为研究对象,采用MTT法检测空白对照组和0.1、0.3、1.0、3.0、10.0、30.0μmol/L黄荆子木脂素3组(n=3)细胞作用48h后的增殖抑制率;采用细胞计数法检测黄荆子木脂素3(10.0μmol/L)组、5-氟尿嘧啶(5-FU,10.0μmol/L)组、溶媒(含0.1%二甲基亚砜的完全培养基)组、空白对照组(n=3)细胞作用24、48、72h后的活细胞数量;采用蛋白印迹法检测溶媒组、黄荆子木脂素3(10.0μmol/L)组、磷脂酰肌醇3-激酶抑制剂LY294002(10.0μmol/L)组及其混合组(n=3)细胞作用24h后的磷酸化(p-)Akt和p-ERK1/2蛋白表达。结果:与空白对照组比较,1.0～30.0μmol/L黄荆子木脂素3组对细胞的抑制率明显升高(P<0.01);与溶媒组和空白对照组比较,黄荆子木脂素3组作用不同时间的活细胞数量均明显减少(P<0.01);与5-FU组比较,黄荆子木脂素3组作用72h后的活细胞数量明显增加(P<0.01);与溶媒组比较,黄荆子木脂素3组、LY294002组及其混合组细胞内p-Akt和p-ERK1/2蛋白表达均明显降低(P<0.01),但后3组间差异无统计学意义。结论:黄荆子木脂素3呈浓度和时间依赖性抑制HepG2细胞的增殖,其机制可能与抑制Akt、ERK1/2蛋白磷酸化水平有关。OBJECTIVE： To study inhibitory effects of Htex negundo lignans-3 on the proliferation of human hepatic cancer HepG2 cells, and relationship of it with Akt and ERK1/2 signal pathway. METHODS： Human hepatic cancer HepG2 cells were as subject, after 48 h treatment the inhibitory rate of cell proliferation was determined by MTT assay in blank control group and 0.1, 0.3, 1.0, 3.0, 10.0 and 30.0 pmol/L Vitex negundo lignans-3 group （n=3）; the viable count of cells were determined by cell count assay in Vitex negundo lignans-3 （10.0 pmol/L） group, 5-FU （10.0 μmol/L） group, solvent （complete medium containing 0.1% DMSO） group and blank control group （n=3） after treated for 24, 48 and 72 h; after 24 h treatment, the expressions of p-Akt and p-ERK1/2 protein were determined by Western blotting assay in solvent group, Htex negundo lignans-3 （10.0 μmol/L） group, phosphatidylinositol-3-kinase blocker LY294002 （10.0 μmol/L） group and mixture group （n=3）. RESULTS： Compared with blank control group, the inhibitory rate of cells increased significantly in 1.0-30.0 μmol/L Htex negundo lignans-3 groups （P〈0.01） ; compared with solvent group and blank control group, the viable count of cells decreased signiftcantly in Vitex negundo lignans-3 group after treated for different time （P〈0.01） ; compared with 5-FU group, the viable count of cells increased significantly in H- tex negundo lignans-3 group after treated for 72 h （P〈0.01）; compared with solvent group, the expressions of p-Akt and p-ERK1/ 2 protien decreased significantly in Fitex negundo lignans-3 group, LY294002 group and mixture group （P〈0.01）, but there was no significant difference among the latter 3 groups. CONCLUSIONS： Vitex negundo lignans-3 inhibits the proliferation of HepG2 cells in concentration and time-dependent manner, associating with the inhibition of Akt and ERK1/2 protein phosphorylation.

关 键 词：人肝癌HEPG2细胞 黄荆子木脂素3 细胞增殖 丝氨酸-苏氨酸蛋白激酶 细胞外调节蛋白激酶1 2 蛋白表达 

分 类 号：R965[医药卫生—药理学]