目的 探讨甘草查尔酮A(LCA)对胶质瘤U87细胞增殖、迁移、侵袭和抗氧化能力的影响及其机制.方法 将体外培养的胶质瘤U87细胞分为4组,空白对照组(常规培养)和低、中、高剂量实验组(5、10、20 μmol·L-1 LCA).用细胞计数试剂盒-8检测细胞增殖能力,用克隆形成实验检测细胞克隆形成能力,用划痕实验法检测细胞迁移能力,用Transwell小室法检测细胞侵袭能力,用比色法检测总谷胱甘肱(T-GSH)、丙二醛(MDA)和超氧化物歧化酶(SOD)水平,用蛋白质印迹法检测磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)的表达水平.结果 空白对照组和低、中、高剂量实验组的细胞增殖活力分别为(90.20±2.17)％、(79.06±1.57)％、(66.13±2.11)％和(49.52±1.82)％,细胞克隆形成率分别为(76.83±2.30)％、(42.33±2.09)％、(17.71±1.84)％和(12.12±1.97)％,12 h 细胞迁移率分别为(34.92±2.24)％、(27.90±1.89)％、(18.76±1.14)％和(14.87±0.82)％,24 h 细胞迁移率分别为(50.37±2.61)％、(39.43±2.56)％、(21.11±2.33)％和(18.32±2.39)％,穿膜细胞数分别为(120.39±4.16)、(79.95±3.83)、(45.67±3.55)和(18.14±2.85)个,T-GSH 水平分别为(71.43±2.39)、(58.51±2.91)、(49.43±2.78)和(35.44±2.76)μmol·L-1,MDA 水平分别为(4.14±0.91)、(7.23±1.75)、(9.20±1.56)和(11.37±1.90)nmol·mL-1,SOD 水平分别为(41.44±2.10)、(35.43±2.91)、(28.56±2.32)和(20.62±2.05)U·mg-1,p-Akt蛋白相对表达水平分别为1.27±0.03、1.06±0.02、0.89±0.01和0.60±0.02.低、中、高剂量实验组的上述指标与空白对照组比较,在统计学上差异均有统计学意义(均P＜0.01).结论 LCA可抑制胶质瘤U87细胞的增殖、迁移、侵袭,诱导氧化损伤,其作用机制可能与下调PI3K/Akt信号通路中p-Akt蛋白的表达相关联.
Effects of Licorice chalcone A on proliferation,migration,invasion and oxidative damage of glioma U87 cells through PI3K/Akt signaling pathway
Objective To investigate the effects of Licorice chalcone A(LCA)on proliferation,migration,invasion and antioxidant capacity of human glioma U87 cells and its mechanism.Methods Glioma U87 cells cultured in vitro were divided into 4 groups,blank control group(conventional culture)and experimental-L,-M,-H groups(5,10,20 μmol·L-1 LC A).Cell proliferation capacity was detected by cell counting kit-8,cell clonogenesis ability was detected by clonogenesis assay,cell migration ability was detected by scratch assay,and cell invasion ability was detected by Transwell assay.Colorimetric assay was used to detect total glutathione(T-GSH),malondialdehyde(MDA)and superoxide dismutase(SOD),and Western blotting was used to detect the protein expression levels of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt).Results The cell proliferation activities of blank control group and experimental-L,-M,-H groups were(90.20±2.17)％,(79.06±1.57)％,(66.13±2.11)％and(49.52±1.82)％;cell clone formation rates were(76.83±2.30)％,(42.33±2.09)％,(17.71±1.84)％and(12.12±1.97)％;12 h cell mobility rates were(34.92±2.24)％,(27.90±1.89)％,(18.76±1.14)％and(14.87±0.82)％;24 h cell mobility rates were(50.37±2.61)％,(39.43±2.56)％,(21.11±2.33)％and(18.32±2.39)％;the number of perforated cells were 120.39±4.16,79.95±3.83,45.67±3.55 and 18.14±2.85;T-GSH levels were(71.43±2.39),(58.51±2.91),(49.43±2.78)and(35.44±2.76)μmol·L-1;MDA levels were(4.14±0.91),(7.23±1.75),(9.20±1.56)and(11.37±1.90)nmol·mL-1;SOD levels were(41.44±2.10),(35.43±2.91),(28.56±2.32)and(20.62±2.05)U·mg-1;the relative expression levels of p-Akt were 1.27±0.03,1.06±0.02,0.89±0.01 and 0.60±0.02,respectively.The above indexes were statistically significant between experimental-L,-M,-H groups and blank control group(all P＜0.01).Conclusion LCA can inhibit the proliferation,migration,invasion and induce oxidative damage of glioma U87 cells,and its mechanism may be related to the down-regulation of p-Akt protein expression in PI3K/Akt signaling pathway.
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