吉林大学学报（医学版）2026，Vol.52Issue(1)：70-80,11.DOI:10.13481/j.1671-587X.20260108
缓激肽B1受体拮抗剂ELN441958通过调节Akt/FoxO3a信号通路对肝癌HepG2细胞增殖的抑制作用
Inhibitory effect of Bradykinin 1 receptor antagonist ELN441958 on proliferation of HepG2 cells by regulating Akt/FoxO3a signaling pathway
摘要
Abstract
Objective:To discuss the inhibitory effect of the bradykinin B1 receptor(B1R)antagonist ELN441958 on the growth of hepatocellular carcinoma HepG2 cells and its regulatory mechanism on the protein kinase B(Akt)/forkhead box O3a(FoxO3a)signaling pathway,and to clarify the anti-tumor effect of the B1R antagonist ELN441958 in liver cancer.Methods:After the HepG2 cells were treated with different doses(0,2.5,5.0,10.0,and 20.0 μmol·L-1)of ELN441958 for 24,48,and 72 h,0 μmol·L-1 ELN441958 was regared as control group.CCK-8 method was used to detect the inhibitory rate of proliferation of the cells in various groups.The HepG2 cells were divided into control group,5.0 μmol·L-1 ELN441958 group,10.0 μmol·L-1 ELN441958 group,and 15.0 μmol·L-1 ELN441958 group;5-ethynyl-2'-deoxyuridine(EdU)staining was used to detect the EdU positive cell rates of the HepG2 cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups and the proportions of the cells at different cell cycles;immunofluorescence method was used to detect the expression level of B1R protein in the HepG2 cells in various groups;Western blotting method was used to detect the expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cyclin D1(Cyclin D1),cyclin-dependent kinase 4(CDK4),B1R,Akt,phosphorylated Akt(p-Akt),FoxO3a,and phosphorylated FoxO3a(p-FoxO3a)proteins in the HepG2 cells in various groups.The HepG2 cells were divided into control group,B1R agonist des-Arg9-BK(1.0 μmol·L-1 des-Arg9-BK)group,and BK+ELN441958 group;Western blotting method was used to detect the expression levels of Bcl-2,Bax,Cyclin D1,CDK4,Akt,p-Akt,FoxO3a,and p-FoxO3a proteins in the HepG2 cells in various groups.The HepG2 cells were divided into control group,Akt agonist SC79 group,and SC79+ELN441958 group;Western blotting method was used to detect the expression levels of Bcl-2,Bax,Cyclin D1,CDK4,Akt,p-Akt,FoxO3a,and p-FoxO3a proteins in the HepG2 cells in various groups.Results:The CCK-8 assay results showed that compared with control group,when the action time was the same,the inhibitory rate of proliferation of the HepG2 cells was significantly increased with the increase of ELN441958 dose(P＜0.05 or P＜0.01);when the dose of ELN441958 was the same,the inhibitory rate of proliferation of the HepG2 cells was significantly increased with the extension of action time(P＜0.05 or P＜0.01).The half maximal inhibitory concentration(IC50)values of ELN441958 at 24,48,and 72 h were(21.4±1.1),(10.5±0.3),and(3.2±0.3)μmol·L-1,respectively.Compared with control group,the EdU positive cell rates of the HepG2 cells in 5.0,10.0,and 15.0 μmol·L-1 ELN441958 groups were significantly decreased(P＜0.05 or P＜0.01).The flow cytometry results showed that compared with control group,the apoptotic rates of the HepG2 cells in different doses of ELN441958 groups were significantly increased(P＜0.05 or P＜0.01),and the percentages of the cells at G0/G1 phase were significantly increased(P＜0.05 or P＜0.01).The Western blotting results showed that compared with control group,the;the expression levels of Bcl-2,CDK4,and Cyclin D1 proteins in the HepG2 cells in 5.0,10.0 and 15.0 μmol·L-1 ELN441958 groups were significantly decreased(P＜0.01),and the expression level of Bax protein was significantly increased(P＜0.05 or P＜0.01).The immunofluorescence results showed that compared with control group,the fluorescence intensity of B1R in the HepG2 cells in 15.0 μmol·L-1 ELN441958 group was significantly decreased(P＜0.01).The Western blotting results showed that compared with control group,the expression level of B1R,p-Akt,and p-FoxO3a proteins in the HepG2 cells in 10.0 and 15.0 μmol·L-1 ELN441958 groups were significantly decreased(P＜0.01);the expressions p-Akt,and p-FoxO3a proteins were decreased(P＜0.05 or P＜0.01)compared with B1R agonist des-Arg9-BK group,the expression levels of Bcl-2,CDK4,Cyclin D1,B1R,p-Akt,and p-FoxO3a proteins in the HepG2 cells in BK+ELN441958 group were significantly decreased(P＜0.01),and the expression level of Bax protein was significantly increased(P＜0.01);compared with SC79 group,the expression levels of Bcl-2,CDK4,Cyclin D1,B1R,p-Akt,and p-FoxO3a proteins in the HepG2 cells in SC79+ELN441958 group were significantly decreased(P＜0.01),and the expression level of Bax protein was significantly increased(P＜0.01).Conclusion:The B1R antagonist ELN441958 can inhibit the proliferation of HepG2 cells,and induce the apoptosis and G0/G1 phase cell cycle arrest of HepG2 cells,and its mechanism may be related to the regulation of Akt/FoxO3a signaling axis.关键词
ELN441958/肝肿瘤/蛋白激酶B/叉头框O3a/细胞凋亡/细胞周期Key words
ELN441958/Liver neoplasms/Protein kinase B/Forkhead box O3a/Apoptosis/Cell cycle分类
医药卫生引用本文复制引用
孙杉杉,陆梅,高新富,吕光耀,赵宝磊,吕文文..缓激肽B1受体拮抗剂ELN441958通过调节Akt/FoxO3a信号通路对肝癌HepG2细胞增殖的抑制作用[J].吉林大学学报（医学版）,2026,52(1):70-80,11.基金项目
山东省科技厅自然科学基金项目(ZR2021MH173) （ZR2021MH173）
山东省卫健委齐鲁卫生与健康杰出青年人才项目(鲁卫人才字[2020]3号) （鲁卫人才字[2020]3号）
