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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 investigate the inhibitory effect of Luteinolus vulgaris 3 on the proliferation of HepG2 human hepatocellular carcinoma cells and its relationship with serine-threonine protein kinase (Akt) and extracellular regulated protein kinase 1/2 (ERK1 / 2) signaling pathway. Methods: HepG2 cells were cultured in HepG2 cells. MTT assay was used to detect the proliferation of blank control group and the cells of three groups (n = 3) (0.1,0.3,1.0,3.0,10.0,30.0μmol / L) for 48h The inhibition rate was measured by using the cell count method. The levels of luteinolignan 3 (10.0μmol / L), 5-fluorouracil (5μmol / L, 10.0μmol / L) (N = 3). The number of viable cells after 24h, 48h and 72h in the blank control group (n = 3) was measured by Western blotting. The levels of viable cells in the media group, 10.0μmol / L rosuvastatin group, - phosphorylated (p-) Akt and p-ERK1 / 2 protein expression in LY294002 (10.0μmol / L) group and its mixed group (n = 3) Results: Compared with the blank control group, the inhibitory rate of 1.0-30.0μmol / L xanthophyllignan 3 group was significantly increased (P <0.01); compared with the vehicle group and the blank control group, (P <0.01). Compared with 5-FU group, the number of viable cells in three groups increased significantly after 72h treatment (P <0.01). Compared with the vehicle group, The expressions of p-Akt and p-ERK1 / 2 in the three groups of lipids and LY294002 group and the mixed group were significantly decreased (P <0.01), but there was no significant difference in the latter three groups. CONCLUSION: Luteintriol 3 inhibits the proliferation of HepG2 cells in a concentration-dependent and time-dependent manner, which may be related to the inhibition of Akt and ERK1 / 2 phosphorylation.