胡桃醌对胃癌细胞耐药性及细胞中ANXA2、ERCC1表达和AKT/mTOR信号通路的影响

Effect of Juglone on cisplatin resistance in gastric cancer cells and expressions of ANXA2,ERCC1 and AKT/m TOR signaling

目的研究胡桃醌对胃癌细胞顺铂耐药性及细胞中膜联蛋白A2(ANXA2)和切除修复交叉互补基因1(ERCC1)表达,及对AKT/mTOR信号通路的影响。方法将BGC-823/DDP细胞株随机分为5组:对照组、胡桃醌组、顺铂组、胡桃醌联合顺铂组、LY294002(AKT抑制剂)联合顺铂组。采用MTT法和流式细胞术检测细胞活性和凋亡率,Western blotting检测细胞中相关蛋白表达。结果与对照组比较,顺铂组细胞凋亡率及细胞中Caspase-3、Caspase-9蛋白表达水平升高(P<0.05),而ANXA2、ERCC1、p-AKT、p-mTOR蛋白表达水平降低(P<0.05);与顺铂组比较,胡桃醌不同浓度组细胞中P-gp、MRP1蛋白表达水平降低(P<0.05),细胞增殖抑制率、细胞凋亡率及细胞中Caspase-3、Caspase-9蛋白表达水平升高(P<0.05),ANXA2、ERCC1、p-AKT、p-mTOR蛋白表达水平降低(P<0.05);与顺铂组比较,LY294002处理后,细胞增殖抑制率和凋亡率升高(P<0.05)。结论胡桃醌增强胃癌细胞对顺铂的敏感性,抑制细胞中ANXA2和ERCC1表达,抑制AKT/mTOR信号通路。

Objective To investigate the effects of Juglone on cisplatin resistance in gastric cancer cells and the expressions of membrane linked protein A2( ANXA2) and excision repair cross complementary gene 1( ERCC1) in cells,as well as the AKT/m TOR signaling pathway.Methods BGC-823/DDP cell lines were randomly divided into 5 groups:normal control group,Juglone group,cisplatin group,Juglone combined with cisplatin group,and LY294002( AKT inhibitor) combined with cisplatin group. MTT assay and flow cytometry were used to detect cell activity and apoptosis rate,and Western blotting was used to detect the expression of related proteins in cells.Results Compared with the control group,the apoptosis rate and the expressions of Caspase-3 and Caspase-9 in the cisplatin group were increased( P<0.05),while the expressions of ANXA2,ERCC1,p-AKT and p-mTOR were decreased( P<0.05). Compared with the cisplatin group,the expressions of P-gp and MRP1 in cells with different concentrations of Juglone were decreased( P<0.05),the proliferation inhibition rate,apoptosis rate,and the expressions of Caspase-3 and Caspase-9 in cells were increased( P<0.05),and the expressions of ANXA2,ERCC1,p-AKT and p-mTOR were decreased( P < 0.05). Compared with cisplatin group,cell proliferation inhibition rate and apoptosis rate increased after LY294002 treatment( P<0.05).Conclusion Juglone enhances the sensitivity of gastric cancer cells to cisplatin,inhibits the expression of ANXA2 and ERCC1 in the cells,and inhibits the AKT/m TOR signaling pathway.