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Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines

  • Authors:
    • Eser Çakmak
    • Birşen Bilgici
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    Affiliations: Department of Medical Biochemistry, Faculty of Medicine, Ondokuz Mayıs University, Atakum, Samsun 55139, Turkey
    Copyright: © Çakmak et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 111
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    Published online on: February 13, 2026
       https://doi.org/10.3892/etm.2026.13106
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Abstract

The antidiabetic drug metformin has potential as an anticancer agent, particularly due to its observed efficacy in breast cancer. Metformin exerts its cytotoxic effects in the induction of endoplasmic reticulum (ER) stress, which can trigger apoptotic cell death pathways. Therefore, the present study aimed to investigate the dose‑dependent effects of metformin on ER stress and apoptosis in HER2‑positive breast cancer SKBR3 cells. For this purpose, SKBR3 cells were treated with 5, 10 and 20 mM metformin. Cell proliferation was assessed using real‑time cell analysis, while expression levels of ER stress‑associated genes [glucose‑regulated protein 78 kDa (GRP78), PRKR‑like ER kinase (PERK), inositol‑requiring enzyme 1 (IRE1), activating transcription factor 6 (ATF6) and CHOP)] were measured by revese transcription‑quantitative PCR. Apoptosis was analyzed by Annexin V‑FITC/PI flow cytometry in cells treated with 10 and 20 mM metformin. Findings revealed that metformin (5, 10 and 20 mM) dose‑dependently inhibited cell proliferation and activated ER stress pathways. Significant increases were observed in gene expression following treatment with 5, 10 and 20 mM metformin, respectively, including  GRP78 (3.70‑, 5.06‑ and 7.33‑fold; all P<0.0001) PERK (2.48‑, 4.36‑ and 9.11‑fold; all P<0.0001), IRE1 (2.15‑fold, P=0.001; 2.90‑fold, P<0.001; 5.55‑fold, P<0.0001), ATF6 (2.43‑2.44‑ and 3.63‑fold; all P<0.0001) and particularly in pro‑apoptotic CHOP (3.31‑, 27.47‑ and 49.85‑fold; all P<0.0001). Flow cytometry revealed that 10 and 20 mM metformin significantly increased early apoptosis to 6.05% (P<0.001) and 7.28% (P<0.001) and late apoptosis to 13.24% (P<0.001) and 20.59% (P<0.001), respectively, compared with controls (early apoptosis, 0.02%; late apoptosis, 0.05%). The present findings demonstrated that metformin activates ER stress response and induces apoptosis in HER2‑positive breast cancer cells in a dose‑dependent manner. This supports the potential of metformin as an adjuvant therapy, though further in vivo studies are needed to evaluate its clinical applicability.
View Figures

Figure 1

Cell proliferation following
metformin treatment. All doses of metformin exhibited
antiproliferative activity.

Figure 2

Gene expression changes of GRP78.
Metformin increased GRP78 gene expression in a dose-dependent
manner ****P<0.0001. GRP78, glucose-regulated protein
78 kDa.

Figure 3

Gene expression changes of PERK.
Metformin increased PERK gene expression in a dose-dependent manner
****P<0.0001. PERK, PRKR-like ER kinase.

Figure 4

Gene expression changes of IRE1.
Metformin increased IRE1 gene expression in a dose-dependent manner
**P<0.01, ***P<0.001 and
****P<0.0001. IRE1, inositol-requiring enzyme 1.

Figure 5

Gene expression changes of ATF6.
Metformin increased ATF6 gene expression
****P<0.0001. ATF6, activating transcription factor
6.

Figure 6

Gene expression changes of CHOP.
Metformin increased CHOP gene expression in a dose-dependent
manner. ****P<0.0001. CHOP, C/EBP homologous
protein.

Figure 7

Flow cytometry analysis of
metformin-induced apoptotic death. Treatment with 10 and 20 mM
metformin significantly increased early apoptotic cells to 6.05 and
7.28 and late apoptotic cells to 13.24 and 20.59% (all P<0.001),
respectively, compared with the control group.

Figure 8

Percentage of early and late
apoptotic cells following metformin treatment. Metformin increased
the percentage of early and late apoptotic cells in a
dose-dependent manner. *P<0.001.
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Copy and paste a formatted citation
Spandidos Publications style
Çakmak E and Bilgici B: Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines. Exp Ther Med 31: 111, 2026.
APA
Çakmak, E., & Bilgici, B. (2026). Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines. Experimental and Therapeutic Medicine, 31, 111. https://doi.org/10.3892/etm.2026.13106
MLA
Çakmak, E., Bilgici, B."Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines". Experimental and Therapeutic Medicine 31.4 (2026): 111.
Chicago
Çakmak, E., Bilgici, B."Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines". Experimental and Therapeutic Medicine 31, no. 4 (2026): 111. https://doi.org/10.3892/etm.2026.13106
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Spandidos Publications style
Çakmak E and Bilgici B: Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines. Exp Ther Med 31: 111, 2026.
APA
Çakmak, E., & Bilgici, B. (2026). Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines. Experimental and Therapeutic Medicine, 31, 111. https://doi.org/10.3892/etm.2026.13106
MLA
Çakmak, E., Bilgici, B."Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines". Experimental and Therapeutic Medicine 31.4 (2026): 111.
Chicago
Çakmak, E., Bilgici, B."Metformin triggers apoptosis via endoplasmic reticulum stress in HER2‑positive breast cancer cell lines". Experimental and Therapeutic Medicine 31, no. 4 (2026): 111. https://doi.org/10.3892/etm.2026.13106
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