紫草素促进食管癌细胞凋亡及细胞周期阻滞的作用机制研究

Effect and mechanism of shikonin on apoptosis and cell cycle arrest of esophageal cancer cells

目的研究紫草素促进食管癌细胞凋亡及细胞周期阻滞的作用及机制。方法2019年1月至2020年6月期间开展细胞实验,培养食管癌Eca109细胞,分为不含药物杜氏改良Eagle培养基(DMEM)处理的对照组,含不同剂量紫草素DMEM处理的紫草素组,含2.0µmol/L紫草素和10µg/L胰岛素样生长因子-1(IGF-1)DMEM处理的2.0µmol/L紫草素+10µg/L IGF-1组。检测细胞凋亡率、细胞周期及凋亡基因活化的胱天蛋白酶-3(cleaved caspase-3)、细胞周期蛋白D1(cyclin D1)、磷酸化磷酸肌醇3激酶(PI3K)、蛋白激酶B(AKT)的表达量。结果0.5µmol/L紫草素组、1.0µmol/L紫草素组、2.0µmol/L紫草素组的细胞凋亡率、细胞周期G1期比例、cleaved caspase-3的表达量[0.5µmol/L紫草素组(0.64±0.14)、1.0µmol/L紫草素组(0.81±0.19)、2.0µmol/L紫草素组(1.02±0.24)]高于对照组0.41±0.07,细胞周期S期、G2期比例及cyclin D1[0.5µmol/L紫草素组(0.80±0.16)、1.0µmol/L紫草素组(0.68±0.13)、2.0µmol/L紫草素组(0.45±0.09)]、p-PI3K、p-AKT表达量低于对照组[cyclin D1表达量(0.91±0.18)](P<0.05);2.0µmol/L紫草素+10µg/L IGF-1组的细胞凋亡率、细胞周期G1期比例、cleaved caspase-3的表达量低于2.0µmol/L紫草素组,细胞周期G2期比例及cyclin D1、p-PI3K、p-AKT表达量高于2.0µmol/L紫草素组(P<0.05)。结论紫草素具有促进食管癌细胞凋亡及细胞周期阻滞的作用,该作用与抑制PI3K/AKT通路激活有关。

Objective To study the effect and mechanism of shikonin on apoptosis and cell cycle arrest of esophageal cancer cells.Methods Cell experiments were carried out from January 2019 to June 2020.Esophageal cancer ECA 109 cells were cultured and assigned into groups:control group treated with Duchenne modified Eagle medium(DMEM)without drug,shikonin groups treated with DMEM containing different doses of shikonin,which included 2.0µmol/L shikonin+10µg/L insulin-like growth factor-1(IGF-1)group treated with DMEM containing 2.0µmol/L shikonin and 10µg/L IGF-1.Apoptosis rate,cell cycle and the expressions of apoptotic gene-activated caspase-3(cleaved caspase-3),cyclin D1,phosphorylated phosphoinositol 3 kinase(PI3K)and protein kinase B(AKT)were detected.Results The apoptotic rate,the proportion of G1 phase and the expression of cleaved caspase-3[0.5µmol/L shikonin group(0.64±0.14),1.0µmol/L shikonin group(0.81±0.19),2.0µmol/L shikonin group(1.02±0.24)]in 0.5µmol/L,1.0µmol/L and 2.0µmol/L shikonin groups were higher than those in the control group 0.41±0.07,while the proportions of S phase and G2 phase and the expressions of cyclin D1[0.5µmol/L shikonin group(0.80±0.16),1.0µmol/L shikonin group(0.68±0.13),2.0µmol/L shikonin group(0.45±0.09)],p-PI3K and p-AKT were lower than those in the control group[cyclin D1 expression:(0.91±0.18)](P<0.05).The apoptotic rate,the proportion of G1 phase and the expression of cleaved caspase-3 in 2.0µmol/L shikonin+10µg/L IGF-1 group were lower than those in 2.0µmol/L shikonin group,while the proportion of G2 phase and the expressions of cyclin D1,p-PI3K and p-AKT were higher than those in 2.0µmol/L shikonin group(P<0.05).Conclusion Shikonin can promote the apoptosis and cell cycle arrest of esophageal cancer cells,which is related to the inhibition of PI3K/Akt pathway activation.