| Zhang Xueyuan,Song Chunyang,Yan Ke,et al.Effect of high mobility group box 1 (HMGB1) protein-mediated radiation bystander effect on the radiosensitivity of esophageal squamous cell carcinoma cells[J].Chinese Journal of Radiological Medicine and Protection,2026,46(1):36-42 |
| Effect of high mobility group box 1 (HMGB1) protein-mediated radiation bystander effect on the radiosensitivity of esophageal squamous cell carcinoma cells |
| Received:February 26, 2025 |
| DOI：10.3760/cma.j.cn112271-20250226-00061 |
| KeyWords:Esophageal Squamous Cell Carcinoma High mobility group box 1 Protein(HMGB1) Radiosensitivity Bystander effect Apoptosis |
| FundProject: |
| Author Name | Affiliation | E-mail | | Zhang Xueyuan | Department of Radiation Therapy, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China | | | Song Chunyang | Department of Radiation Therapy, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China | | | Yan Ke | Department of Radiation Therapy, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China | hbmuzhusc1965@hebmu.edu.cn | | Shen Wenbin | Department of Radiation Therapy, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China | | | Zhu Shuchai | Department of Radiation Therapy, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China | |
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| Abstract:: |
| Objective To investigate the release of high mobility group box 1 (HMGB1) induced by radiation in esophageal squamous cell carcinoma (ESCC) cells and its role in regulating radiosensitivity. Methods Human ESCC cell lines ECA109 and TE13 were used. The concentration of HMGB1 in the cell supernatant was measured by enzyme-linked immunosorbent assay (ELISA) after different doses of X-ray irradiation (0, 2, 4, 6, 8, and 10 Gy) and at different time points (0-24 h) after 10 Gy irradiation. HMGB1-knockdown cell lines were established using shRNA. Supernatants collected from cells 12 h after 10 Gy irradiation (S-IR group) or from unirradiated cells (S group) were applied to the corresponding ECA109 and TE13 cells. Cell proliferation, invasion, apoptosis, and radiosensitivity were evaluated using the CCK-8 assay, Transwell assay, flow cytometry, and clonogenic assay, respectively. Results ELISA results showed that HMGB1 release (ECA109: 49.04-2,098.00 pg/ml; TE13: 47.85-1,883.00 pg/ml) was positively correlated with irradiation dose (peaking at 10 Gy). Moreover, HMGB1 concentrations in the supernatant (ECA109: 49.04-2,128.00 pg/ml; TE13: 47.85-1,983.00 pg/ml) were significantly increased at 12-24 h after irradiation. The CCK-8 assay revealed enhanced proliferative activity in the S-IR-negative control group, while no significant difference was observed in the S-IR-HMGB1-shRNA group compared to the S-control group (P> 0.05). In the Transwell assay, the number of transmigrated cells in the S-IR-negative control group (ECA109: 37.67 ± 3.06; TE13: 68.33 ± 3.51) was significantly higher than that in the S-negative control group (ECA109: 18.33 ± 2.08; TE13: 47.00 ± 6.23) (t = 9.06, 5.16, P< 0.01). HMGB1 knockdown significantly reduced the number of transmigrated cells (t = 5.14, 5.76, P< 0.01). Flow cytometry showed that the apoptosis rate in the S-IR-negative control group was lower than that in the S-IR-HMGB1-shRNA group (t = 4.27, 4.18, P< 0.05). The clonogenic assay demonstrated that, compared with the S-IR-negative control group, the S-IR-HMGB1-shRNA group exhibited significantly reduced values for the mean lethal dose (D0), extrapolation number (N), quasi-threshold dose (Dq), and surviving fraction at 2 Gy (SF2) (ECA109:t = 3.85、8.78、3.81、3.51,P<0.05;TE13:t = 2.81、4.49、2.82、3.32,P<0.05). Conclusions Radiation-induced release of HMGB1 promotes proliferation and invasion, inhibits apoptosis, and reduces the radiosensitivity of ESCC cells. HMGB1 may be a key regulator of the radiation-induced bystander effect. |
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