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. 2021 Jan;23(1):88.
doi: 10.3892/mmr.2020.11725. Epub 2020 Nov 25.

Metformin induces apoptosis and inhibits migration by activating the AMPK/p53 axis and suppressing PI3K/AKT signaling in human cervical cancer cells

Ya-Hui Chen  1 , Shun-Fa Yang  1 , Chueh-Ko Yang  2 , Horng-Der Tsai  3 , Tze-Ho Chen  3 , Ming-Chih Chou  1 , Yi-Hsuan Hsiao  1
Affiliations

Metformin induces apoptosis and inhibits migration by activating the AMPK/p53 axis and suppressing PI3K/AKT signaling in human cervical cancer cells

Ya-Hui Chen et al. Mol Med Rep. 2021 Jan.

Abstract

Human cervical cancer is the fourth most common malignancy among women worldwide, and it is expected to result in 460,000 deaths per year by 2040. Moreover, patients with cervical cancer often display drug resistance and severe side effects; therefore, the development of effective novel chemotherapeutic agents is important. In the present study, the effects of metformin, a first‑line therapeutic drug for type 2 diabetes mellitus, were evaluated in cervical cancer. Compared with the control group, metformin significantly inhibited cell viability and migration, and induced apoptosis and cell cycle arrest in human cervical cancer cell lines (CaSki and HeLa). Following metformin treatment, the protein expression levels of p‑AMP‑activated protein kinase (p‑AMPK), which promotes cell death, and the tumor suppressor protein p‑p53 were remarkably upregulated in CaSki and C33A cells compared with the control group. Furthermore, compared with the control group, metformin significantly suppressed the PI3K/AKT signaling pathway in CaSki, C33A and HeLa cells. Compound C (an AMPK inhibitor) significantly reversed the effects of metformin on CaSki, C33A and HeLa cell viability, and AMPK and p53 phosphorylation. The results of the present study suggested that metformin induced AMPK‑mediated apoptosis, thus metformin may serve as a chemotherapeutic agent for human cervical cancer.

Keywords: metformin; apoptosis; migration; AMPK‑activated protein kinase/p53; PI3K/AKT; cervical cancer.

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Figures

Figure 1.
Figure 1.
Metformin inhibits cervical cancer cell proliferation and migration. (A) CaSki, C33A and HeLa cells were treated with metformin (0–20 mM) for 48 h. Cell viability was determined by performing the Cell Counting Kit-8 assay. (B) CaSki, C33A and HeLa cells were treated with metformin (0–10 mM) for 48 h. Cell migration was assessed by performing a Transwell migration assay (scale bar, 200 µm). Data are presented as the mean ± SD from three independent experiments. *P<0.05 vs. CON. CON, 0 mM metformin; M5, 5 mM metformin; M10, 10 mM metformin.
Figure 2.
Figure 2.
Metformin induces cervical cancer cell apoptosis. CaSki, C33A and HeLa cells were treated with metformin (0, 5 or 10 mM) for 48 h. Cell apoptosis was (A) determined via flow cytometry and (B) quantified. Data are presented as the mean ± SD from three independent experiments. *P<0.05 vs. CON. CON, 0 mM metformin; M5, 5 mM metformin; M10, 10 mM metformin.
Figure 3.
Figure 3.
Metformin induces cervical cancer cell cycle arrest. CaSki, C33A and HeLa cells were treated with metformin (0, 5 or 10 mM) for 48 h. Cell cycle phase distribution was (A) determined via flow cytometry and (B) quantified. Data are presented as the mean ± SD from three independent experiments. *P<0.05 vs. CON. CON, 0 mM metformin; M5, 5 mM metformin; M10, 10 mM metformin.
Figure 4.
Figure 4.
Effect of metformin on AMPK and caspase-dependent apoptosis signaling in cervical cancer cell lines. CaSki, C33A and HeLa cells were treated with metformin (0, 5 or 10 mM) for 48 h. p-AMPK, AMPK, p-p53, p53, Bcl-2, Bak, Bax and cleaved caspase-3 protein expression levels were (A) determined via western blotting and semi-quantified in (B) CaSki, (C) C33A and (D) HeLa cells. Data are presented as the mean ± SD from three independent experiments. *P<0.05 vs. CON. AMPK, AMP-activated protein kinase; p, phosphorylated; Bak, Bcl-2 antagonist/killer 1; CON, 0 mM metformin; M5, 5 mM metformin; M10, 10 mM metformin; C-cas-3, cleaved caspase-3.
Figure 5.
Figure 5.
Effects of metformin on PI3K/AKT/mTOR signaling in cervical cancer cell lines. CaSki, C33A and HeLa cells were treated with metformin (0, 5 or 10 mM) for 48 h. Protein expression levels of PIK3CA, p-AKT, AKT, p-p70S6K, and p70S6K were (A) determined via western blotting and semi-quantified in (B) CaSki, (C) C33A and (D) HeLa cells. Data are presented as the mean ± SD from three independent experiments. *P<0.05 vs. CON. p, phosphorylated; PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α; p70S6K, p70S6 kinase; CON, 0 mM metformin; M5, 5 mM metformin; M10, 10 mM metformin.
Figure 6.
Figure 6.
Effects of Compound C on cell viability, AMPK signaling and apoptotic signaling in cervical cancer cell lines. CaSki, C33A and HeLa cells were pre-treated with or without Compound C (1 or 5 µM; an AMPK inhibitor) for 2 h and then treated with or without 10 mM metformin for 48 h, the concentration of metformin selected for experiment was based on the cell viability assay test. (A) Cell viability was determined by performing the Cell Counting Kit-8 assay. Protein expression levels of p-AMPK, AMPK, p-p53, p53, Bcl-2 and cleaved caspase-3 were (B) determined by western blotting and semi-quantified in (C) CaSki, (D) C33A and (E) HeLa cells. Data are presented as the mean ± SD from three independent experiments. *P<0.05 vs. CON; †P<0.05 vs. M10. AMPK, AMP-activated protein kinase; p, phosphorylated; CON, 0 mM metformin; M10, 10 mM metformin; Com C1, 1 µM Compound C; Com C5, 5 µM Compound C; C-cas-3, cleaved caspase-3.
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References

    1. Small W, Jr, Bacon MA, Bajaj A, Chuang LT, Fisher BJ, Harkenrider MM, Jhingran A, Kitchener HC, Mileshkin LR, Viswanathan AN, Gaffney DK. Cervical cancer: A global health crisis. Cancer. 2017;123:2404–2412. doi: 10.1002/cncr.30667. - DOI - PubMed
    1. Simms KT, Steinberg J, Caruana M, Smith MA, Lew JB, Soerjomataram I, Castle PE, Bray F, Canfell K. Impact of scaled up human papillomavirus vaccination and cervical screening and the potential for global elimination of cervical cancer in 181 countries, 2020-99: A modelling study. Lancet Oncol. 2019;20:394–407. doi: 10.1016/S1470-2045(18)30836-2. - DOI - PubMed
    1. Ch PN, Gurram L, Chopra S, Mahantshetty U. The management of locally advanced cervical cancer. Curr Opin Oncol. 2018;30:323–329. doi: 10.1097/CCO.0000000000000471. - DOI - PubMed
    1. Rosen VM, Guerra I, McCormack M, Nogueira-Rodrigues A, Sasse A, Munk VC, Shang A. Systematic review and network meta-analysis of bevacizumab plus first-line topotecan-paclitaxel or cisplatin-paclitaxel versus non-bevacizumab-containing therapies in persistent, recurrent, or metastatic cervical cancer. Int J Gynecol Cancer. 2017;27:1237–1246. doi: 10.1097/IGC.0000000000001000. - DOI - PMC - PubMed
    1. Cervical Cancer. National Cancer Institute; May, 2020. Bethesda. SEER Cancer Stat Facts.

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