艾迪注射液对人胃癌HGC-27细胞的抗肿瘤作用机制研究

Anti-tumor mechanism of Aidi injection on human gastric cancer HGC 27 cells

目的探讨艾迪注射液对人胃癌HGC-27细胞增殖和迁移能力的影响及作用机制研究。方法采用0、7.5、15.0、30.0、60.0、120.0 mg/ml艾迪注射液处理人胃癌HGC-27细胞,采用噻唑蓝(MTT)法检测细胞增殖能力。不同浓度艾迪注射液培养人胃癌HGC-27细胞48 h的半数抑制浓度(IC50)为20.17 mg/ml,后续实验将6.0、12.0、24.0 mg/ml艾迪注射液处理的人胃癌HGC-27细胞设置为低浓度组、中浓度组、高浓度组,以野生型HGC-27细胞作为空白对照组,2.0 mg/L顺铂处理的胃癌细胞HGC-27作为阳性对照组,细胞划痕试验检测细胞的迁移能力,蛋白质印迹法(Westernblot)检测各组细胞蛋白激酶B(AKT)、雷帕霉素靶蛋白(MTOR)、真核起始因子4E结合蛋白1(4EBP1)、磷酸化的4EBP1(p-4EBP1)蛋白的相对表达量。结果随艾迪注射液浓度的升高,培养24、48、72 h,人胃癌HGC-27细胞A值均逐渐降低,差异均有统计学意义(P﹤0.05)。培养24 h,低浓度组、中浓度组、高浓度组人胃癌HGC-27细胞未迁移率均高于空白对照组(P﹤0.05),高浓度组人胃癌HGC-27细胞未迁移率高于低浓度和中浓度组(P﹤0.05)。低浓度组和中浓度组人胃癌HGC-27细胞AKT、4EBP1、p-4EBP1蛋白的相对表达量均低于空白对照组(P﹤0.05),且中浓度组人胃癌HGC-27细胞MTOR蛋白相对表达量低于空白对照组(P﹤0.05),高浓度组人胃癌HGC-27细胞AKT、MTOR、4EBP1、p-4EBP1蛋白相对表达量均低于低浓度组和中浓度组,中浓度组人胃癌HGC-27细胞AKT蛋白相对表达量高于低浓度组,差异均有统计学意义(P﹤0.05)。结论艾迪注射液能够抑制人胃癌HGC-27细胞的增殖和迁移能力,且MTOR信号通路可能是其抗胃癌治疗的关键。

Objective To investigate the effect of Aidi injection on the proliferation and migration of human gastric cancer HGC-27 cells and its underlying mechanism.Method Human gastric cancer HGC-27 cells were treated with 0,7.5,15.0,30.0,60.0,120.0 mg/ml Aidi injection,and the cell proliferation ability was detected by the methyl thiazolyl tetrazolium(MTT)method.The half inhibit concentration(IC50)of Aidi injection on human gastric cancer HGC-27 cells for48 hours was 20.17 mg/ml.The subsequent experiments used 6.0,12.0,and 24.0 mg/ml of Aidi injection for human gastric cancer HGC-27 cells as low concentration group,medium concentration group,high concentration group,with wildtype HGC-27 cells as a blank control group,and gastric cancer HGC-27 cells treated with 2.0 mg/L cisplatin as a positive control group.The detection of cell migration was based on the cell scratch method.The relative expression of protein kinase B(AKT),mechanistic target of rapamycin kinase(MTOR),eukaryotic translation initiation factor 4 E binding protein 1(4 EBP1),and phospho-4 EBP1(p-4 EBP1)protein were measured by the Western blot in each group.Result With the increase of the concentration of Aidi injection,the A value of human gastric cancer HGC-27 cells gradually decreased after 24,48,and 72 hours of culture,and the differences were statistically significant(P<0.05).After 24 hours of culture,the non-migration rate of human gastric cancer HGC-27 cells in the low concentration group,medium concentration group,and high concentration group was higher than that of the blank control group(P<0.05),and the non-migration rate of human gastric cancer HGC-27 cells in the high concentration group was higher than that in the low concentration and medium concentration group(P<0.05).The relative expression levels of AKT,4 EBP1,and p-4 EBP1 in human gastric cancer HGC-27 cells in the low concentration group and the medium concentration group were lower than those in the blank control group(P<0.05),and the relative expression of MTOR protein in human gastric cancer HGC-27 cells in the medium concentration group was lower than that in the blank control group(P<0.05).The relative expression levels of AKT,MTOR,4 EBP1,p-4 EBP1 protein in human gastric cancer HGC-27 cells in the high concentration group were lower than those in the low concentration group and the medium concentration group,and the relative expression of AKT protein in the medium concentration group was higher than that in the low concentration group,and the differences were statistically significant(P<0.05).Conclusion Aidi injection can inhibit the proliferation and migration of human gastric cancer HGC-27 cells,and the MTOR signal pathway may be the key to its anti-gastric cancer treatment.