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Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway

  • Authors:
    • Yijun Wang
    • Mengyang Zhao
    • Shanshan Liu
    • Rui Zheng
    • Tianhui Gao
  • View Affiliations / Copyright

    Affiliations: Department of Oncology, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Zhengzhou, Henan 450001, P.R. China, State Key Laboratory of Chemical Oncogenomics, Guangdong Provincial Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518071, P.R. China
    Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 38
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    Published online on: November 11, 2025
       https://doi.org/10.3892/mmr.2025.13748
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Abstract

Pancreatic cancer is characterized by short survival and poor treatment outcomes. Astragaloside IV (AST‑IV), the primary pharmacological component of Astragalus membranaceus, is a traditional Chinese medicinal component with demonstrated anticancer potential. The present study aimed to evaluate the therapeutic efficacy of AST‑IV against pancreatic cancer cells in vitro and to elucidate its underlying mechanisms of action, thereby providing novel insights for its clinical application in the treatment of pancreatic cancer. The effects of AST‑IV on PANC‑1 pancreatic cancer cell viability and migration were assessed using Cell Counting Kit‑8 and wound healing assays, respectively. Subsequently, RNA‑sequencing (RNA‑seq) analysis was performed, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to investigate the mechanisms underlying the effects of AST‑IV. Finally, western blotting experiments were conducted to validate the potential molecular mechanisms of AST‑IV. The results revealed that AST‑IV effectively suppressed the proliferation and migration of the pancreatic cancer cells. In addition, GO and KEGG analyses of the differentially expressed genes identified by RNA‑seq analysis suggested that AST‑IV induced endoplasmic reticulum (ER) stress and influenced critical cellular processes, including cell cycle regulation and DNA damage repair. Furthermore, western blotting demonstrated that AST‑IV significantly activated the protein kinase R‑like endoplasmic reticulum kinase (PERK) signaling pathway, upregulated activating transcription factor 4 expression and induced the overexpression of CCAAT/enhancer‑binding protein homologous protein, indicating that it triggered apoptosis. In summary, these findings suggest that AST‑IV induced apoptosis in pancreatic cancer cells through PERK‑mediated ER stress. These results expand the potential therapeutic applications of AST‑IV and provide a theoretical foundation for the development of novel treatment strategies and therapeutic targets for pancreatic cancer treatment.
View Figures

Figure 1

AST-IV inhibits the proliferation and
migration of PANC-1 pancreatic cancer cells. (A) Chemical structure
of AST IV. (B) Viability of PANC-1 cells detected via Cell Counting
Kit-8 assay after treatment for 24, 48 and 72 h with various
concentrations of AST-IV. A growth curve was also constructed after
48 h of AST-IV treatment. (C) Migration of PANC-1 cells detected by
wound healing assay after treatment with 80 µM AST-IV for 24, 48
and 72 h. Scale bar, 100 µm. *P<0.05 and **P<0.01 vs. the
control group. AST-IV, astragaloside IV.

Figure 2

Volcano plot and heatmap of
significant DEGs between the AST-IV treatment and control groups.
(A) Volcano plot of the DEGs shows 1,906 transcripts that were
significantly altered, with 944 upregulated (red) and 962
downregulated (blue) in the AST-IV group. (B) The DEGs were
subjected to cluster analysis and the results are shown as
heatmaps. AST-IV, astragaloside IV; DEGs, differentially expressed
genes; FC, fold change; nodiff, no significant difference.

Figure 3

GO and KEGG enrichment analyses of
upregulated DEGs in the AST-IV group compared with the control
group. (A) GO enrichment of upregulated DEGs following treatment
with 80 µM AST-IV for 48 h. (B) KEGG pathway enrichment analysis of
upregulated DEGs under the same treatment conditions. AST-IV,
astragaloside IV; DEG, differentially expressed genes; GO, Gene
Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Figure 4

GO and KEGG enrichment analysis of
downregulated DEGs in the AST-IV group compared with the control
group. (A) GO enrichment of downregulated DEGs following treatment
with 80 µM AST-IV for 48 h. (B) KEGG pathway enrichment analysis of
downregulated DEGs under the same treatment conditions. AST-IV,
astragaloside IV; DEGs, differentially expressed genes; GO, Gene
Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Figure 5

Analysis of the mechanism of AST-IV
in the treatment of pancreatic cancer. Heatmap of differentially
expressed marker genes involved in (A) ER stress and (B) the cell
cycle. AST-IV, astragaloside IV; ER, endoplasmic reticulum;
lg(FPKM), log10 (fragments per kilobase of transcript
per million mapped fragments).

Figure 6

AST-IV activates the PERK signaling
pathway and induces the apoptosis of pancreatic cancer cells. (A)
Representative western blots of GRP78, PERK, p-PERK, eIF2α and
p-eIF2α and (B) semi-quantitative analysis of the blots. (C)
Representative western blots of ATF4, CHOP, Bcl-2 and Bax and (D)
semi-quantitative analysis of the blots. Data are presented as the
mean ± SEM (n=3). *P<0.05 and **P<0.01 as indicated. AST-IV,
astragaloside IV; eIF2a, eukaryotic translation initiation factor 2
a; ATF4, activating transcription factor 4; CHOP,
CCAAT/enhancer-binding protein homologous protein; GRP78, 78-kDa
glucose-regulated protein; p-, phosphorylated; PERK, protein kinase
R-like endoplasmic reticulum kinase.

Figure 7

Schematic model illustrating the
proposed mechanism by which AST-IV promotes apoptosis in pancreatic
cancer cells. AST-IV induces ER stress via activation of the PERK
signaling pathway, leading to upregulation of the downstream
signaling factor ATF4. This subsequently upregulates the expression
of pro-apoptotic proteins CHOP and Bax, while downregulating the
anti-apoptotic protein Bcl-2, thereby promoting the apoptosis of
pancreatic cancer cells. AST-IV, astragaloside IV; ATF, activating
transcription factor; CHOP, CCAAT/enhancer-binding protein
homologous protein; eIF2a, eukaryotic translation initiation factor
2 a; ER, endoplasmic reticulum; GADD34, growth arrest and DNA
damage-inducible protein 34; GRP78, 78-kDa glucose-regulated
protein; PERK, protein kinase R-like endoplasmic reticulum kinase;
SP1/2, site-1/2 protease; UPR, unfolded protein response.
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Spandidos Publications style
Wang Y, Zhao M, Liu S, Zheng R and Gao T: Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway. Mol Med Rep 33: 38, 2026.
APA
Wang, Y., Zhao, M., Liu, S., Zheng, R., & Gao, T. (2026). Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway. Molecular Medicine Reports, 33, 38. https://doi.org/10.3892/mmr.2025.13748
MLA
Wang, Y., Zhao, M., Liu, S., Zheng, R., Gao, T."Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway". Molecular Medicine Reports 33.1 (2026): 38.
Chicago
Wang, Y., Zhao, M., Liu, S., Zheng, R., Gao, T."Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway". Molecular Medicine Reports 33, no. 1 (2026): 38. https://doi.org/10.3892/mmr.2025.13748
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Spandidos Publications style
Wang Y, Zhao M, Liu S, Zheng R and Gao T: Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway. Mol Med Rep 33: 38, 2026.
APA
Wang, Y., Zhao, M., Liu, S., Zheng, R., & Gao, T. (2026). Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway. Molecular Medicine Reports, 33, 38. https://doi.org/10.3892/mmr.2025.13748
MLA
Wang, Y., Zhao, M., Liu, S., Zheng, R., Gao, T."Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway". Molecular Medicine Reports 33.1 (2026): 38.
Chicago
Wang, Y., Zhao, M., Liu, S., Zheng, R., Gao, T."Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway". Molecular Medicine Reports 33, no. 1 (2026): 38. https://doi.org/10.3892/mmr.2025.13748
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