【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。更多还原

【Abstract】 OBJECTIVE:To study inhibitory effects of Vitex 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 μmol/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 μmol/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,Vitex 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 μ mol/L Vitex negundo lignans-3 groups(P<0.01);compared with solvent group and blank control group,the viable count of cells decreased significantly 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 Vitex 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 Vitex 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.更多还原