Journal of Jilin University(Medicine Edition) ›› 2020, Vol. 46 ›› Issue (6): 1162-1168.doi: 10.13481/j.1671-587x.20200609

• Research in basic medicine • Previous Articles     Next Articles

Effect of astragalus polysaccharide on cisplatin resistance of human lung cancer A549/DDP cells and its mechanism

Ye LIU1,Longyun CHEN2()   

  1. 1.Department of Clinical Microbiology,School of Laboratory Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China
    2.Department of Biochemistry,School of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan 430065,China
  • Received:2020-05-06 Online:2020-11-28 Published:2022-08-24
  • Contact: Longyun CHEN E-mail:bcly988@hbtcm.edu.cn

Abstract: Objective

To explore the effect of astragalus polysaccharide (APS) on the cisplatin(DDP) resistance of human lung cancer A549/DDP cells, and to clarify its possible mechanism.

Methods

The A549/DDP cells at logarithmic growth phase were divided into control group (without drug intervention), APS groups ( given 25, 50, 100, 200, and 400 mg·L-1 APS), DDP groups (given 2, 4, 8, 16, and 32 mg·L-1 DDP) and APS+DDP groups (given 100 mg·L-1 APS and 2, 4, 8, 16, 32 mg·L-1 DDP). After the cells were treated for 24 h, the inhibitory rates of proliferation of the A549/DDP cells in various groups were detected by CCK-8 method and the half inhibitory concentration (IC50) was caluculated. The A549/DDP cells were divided into control group (without drug intervention), APS group (given 100 mg·L-1 APS), DDP group (given 11.46 mg·L-1 DDP) and APS+DDP group (given 100 mg·L-1 APS and 11.46 mg·L-1 DDP), the cells in various groups were treated with drugs for 24 h. The migration of A549/DDP cells in various groups was detected by Transwell chamber assay; the apoptotic rates and mitochondrial membrane potentials of the A549/DDP cells in various groups were detected by flow cytometry, and the level expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax),P-glycoprotein (P-gp),phosphatidylinositol-3 kinase (PI3K), phosphorylated phosphatidylinositol-3 kinase (p-PI3K), protein kinase B (AKT) and phosphorylated protein kinase B (p-AKT) proteins in the A549 / DDP cells in various groups were detected by Western blotting method.

Results

The CCK-8 results showed that compared with DDP group, the IC50 of DDP in APS+DDP group was decreased. The Transwell chamber assay results showed that compared with control group, the migration number of A549 / DDP cells in APS group was not significantly changed; compared with DDP group, the migration number of A549 / DDP cells in APS+ DDP group was decreased.The results of flow cytometry showed that compared with control group, the apoptotic rate and mitochondrial membrane potential of A549 / DDP cells in APS group had no significant changes (P>0.05), and the apoptotic rate and mitochondrial membrane potential of the A549/DDP cells in DDP group were significantly increased (P<0.05); compared with DDP group, the apoptotic rate and mitochondrial membrane potential of A549 / DDP cells in APS+DDP group were significantly increased (P<0.05);the Western blotting results showed that compared with control group, the expression levels of Bcl-2 and P-gp proteins in the A549/DDP cells in APS group was significantly decreased (P<0.05), the ratios of p-PI3K/PI3K and p-AKT/AKT were decreased(P<0.05), and there was no significant difference in the expression level of Bax protein (P>0.05);compared with APS group, the expression levels of Bcl-2 and P-gp proteins in the A549 / DDP cells in DDP group were significantly decreased (P<0.05),the ratios of p-PI3K/PI3K and p-AKT/AKT were decreased(P<0.05), and the expression level of Bax protein was significantly increased (P<0.05);compared with DDP group, the expression levels of Bcl-2 and P-gp proteins in the A549 / DDP cells in DDP+APS group were significantly decreased (P<0.05), the ratios of p-PI3K/PI3K and p-AKT/AKT were decreased(P<0.05),and the expression level of Bax protein was significantly increased (P<0.05).

Conclusion

APS can reverse the drug resistance of A549/DDP cells,and its mechanism may be related to blocking the PI3K/AKT signal pathway and inducing the mitochondrial damage.

Key words: astragalus polysaccharide, A549/DDP cells, drug resistance, phosphatidylinositol-3 kinase, protein kinase B

CLC Number: 

  • R273