Metformin suppresses esophageal cancer progression through the radiation‑induced cellular senescence of cancer‑associated fibroblasts

Sugimoto,Yuya; Okamoto,Koichi; Saito,Hiroto; Yamaguchi,Takahisa; Kinoshita,Jun; Nakamura,Keishi; Takino,Takahisa; Endo,Yoshio; Ninomiya,Itasu; Ohta,Tetsuo; Inaki,Noriyuki

doi:10.3892/or.2024.8788

Metformin suppresses esophageal cancer progression through the radiation‑induced cellular senescence of cancer‑associated fibroblasts

Authors:
- Yuya Sugimoto
- Koichi Okamoto
- Hiroto Saito
- Takahisa Yamaguchi
- Jun Kinoshita
- Keishi Nakamura
- Takahisa Takino
- Yoshio Endo
- Itasu Ninomiya
- Tetsuo Ohta
- Noriyuki Inaki
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Affiliations: Department of Gastrointestinal Surgery, Division of Cancer Medicine, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920‑8641, Japan, Department of Gastroenterological Surgery, Ishikawa Prefectural Central Hospital, Kanazawa, Ishikawa 920‑8530, Japan, Division of Education for Global Standard, Institute of Liberal Arts and Science, Kanazawa University, Kanazawa, Ishikawa 920‑1192, Japan, Central Research Resource Branch, Cancer Research Institute, Kanazawa University, Kanazawa, Ishikawa 920‑1192, Japan, Department of Surgery, Fukui Prefectural Hospital, Fukui 910‑0846, Japan
Published online on: August 2, 2024 https://doi.org/10.3892/or.2024.8788
Article Number: 129
Copyright: © Sugimoto et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Senescent cells are known to secrete proteins, including inflammatory cytokines and damage‑associated molecular patterns. This phenomenon is known as the senescence‑associated secretory phenotype (SASP). SASP in cancer stromal fibroblasts is involved in cancer growth and progression. Conversely, metformin, an antidiabetic drug, has been reported to inhibit SASP induction by inhibiting the activation of NF‑κB, a regulator of SASP. To date, at least to the best of our knowledge, there have been no reports regarding cellular senescence in fibroblasts and tumor progression via the SASP‑mediated paracrine pathway. The present study thus aimed to elucidate the induction mechanisms of SASP in radiation‑induced fibroblasts and to determine its effects on cancer progression via the paracrine pathway. Furthermore, the present study aimed to determine whether controlling SASP using metformin suppresses cancer progression. A well‑differentiated esophageal cancer cell line established by the authors' department and fibroblasts isolated and cultured from the non‑cancerous esophageal mucosa of resected esophageal cancer cases were used for the experiments. Fibroblasts were irradiated with 8 Gy radiation, and the changes in the expression of the senescence markers, SA‑β‑gal, p21, p16 and NF‑κB were evaluated using immunofluorescent staining and western blot analysis in the presence or absence of metformin treatment. The culture supernatants of irradiated fibroblasts treated with metformin and those treated without metformin were collected and added to the cancer cells to evaluate their proliferative, invasive and migratory abilities. Vimentin and E‑cadherin expression levels were also evaluated using immunofluorescent staining and western blot analysis. The expression levels of p16, p21 and NF‑κB in irradiated fibroblasts were attenuated by treatment with metformin. Supernatants collected from irradiated fibroblasts exhibited the proliferative activity of esophageal cancer cells, and the promotion of migratory and invasion abilities, which may be due to epithelial‑mesenchymal transition and changes in cell morphology. These reactions were confirmed to be suppressed by the addition of the supernatant of cultured fibroblasts pre‑treated with metformin. On the whole, the present study demonstrates that fibroblasts in the cancer stroma may be involved in tumor progression through cellular senescence.

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