中华放射医学与防护杂志

张雪原,宋春洋,闫可,等.高迁移率族蛋白B1介导的辐射旁效应对食管鳞癌细胞放射敏感性的影响[J].中华放射医学与防护杂志,2026,46(1):36-42.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].Chin J Radiol Med Prot,2026,46(1):36-42

高迁移率族蛋白B1介导的辐射旁效应对食管鳞癌细胞放射敏感性的影响

Effect of high mobility group box 1 (HMGB1) protein-mediated radiation bystander effect on the radiosensitivity of esophageal squamous cell carcinoma cells

投稿时间：2025-02-26

DOI：10.3760/cma.j.cn112271-20250226-00061

中文关键词: 食管鳞癌高迁移率族蛋白B1 放射敏感性旁效应细胞凋亡

英文关键词:Esophageal Squamous Cell Carcinoma High mobility group box 1 Protein(HMGB1) Radiosensitivity Bystander effect Apoptosis

基金项目:

作者	单位	E-mail
张雪原	河北医科大学第四医院放疗科, 石家庄 050011
宋春洋	河北医科大学第四医院放疗科, 石家庄 050011
闫可	河北医科大学第四医院放疗科, 石家庄 050011	hbmuzhusc1965@hebmu.edu.cn
沈文斌	河北医科大学第四医院放疗科, 石家庄 050011
祝淑钗	河北医科大学第四医院放疗科, 石家庄 050011

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中文摘要:

目的探索辐射诱导食管鳞癌细胞释放高迁移率族蛋白B1(HMGB1)及其对细胞放射敏感性的调控作用。方法选用人食管鳞癌细胞系ECA109和TE13,采用酶联免疫吸附(ELISA)法检测不同剂量X射线(0、2、4、6、8和10 Gy)照射后及10 Gy照射后的不同时间点(0~24 h)细胞上清液中HMGB1的浓度。构建HMGB1-shRNA低表达细胞株,分别收集10 Gy照射后12 h(S-IR组)及未照射(S组)细胞上清处理相应细胞,采用CCK-8法、Transwell实验、流式细胞术、克隆形成实验分别评估细胞增殖、侵袭、调亡和放射敏感性。结果 ELISA检测显示,不同剂量照射后,HMGB1释放量(ECA109:49.04~2 098.00 pg/ml,TE13:47.85~1 883.00 pg/ml)与照射剂量呈正相关(10 Gy组达最高值);且在照射后12~24 h上清HMGB1浓度(ECA109:49.04~2 128.00 pg/ml,TE13:47.85~1 983.00 pg/ml)显著升高。CCK-8实验显示,S-IR-阴性对照组增殖活性增强,而S-IR-HMGB1-shRNA组增殖活性与S-对照组相比差异无统计学意义(P>0.05)。Transwell实验中,S-IR-阴性对照组穿膜细胞数(ECA109,37.67±3.06;TE13,68.33±3.51)显著高于S-阴性对照组(18.33±2.08;47.00±6.23)(t=9.06、5.16,P＜0.01);而HMGB1敲低后穿膜数显著减少(t=5.14、5.76,P＜0.01)。流式细胞术显示,S-IR-阴性对照组凋亡率低于S-IR-HMGB1-shRNA组(t=4.27、4.18,P<0.05)。克隆实验显示,与S-IR-阴性对照组相比,S-IR-HMGB1-shRNA组平均致死剂量(D₀)、外推数(N)、准阈剂量(D_q)及2 Gy射线照射后细胞存活分数(SF₂)值均显著降低,差异均有统计学意义(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)。结论辐射诱导释放的HMGB1通过促进细胞增殖和侵袭、抑制凋亡,降低食管鳞癌细胞放射敏感性,可能是辐射旁效应的关键调控因子。

英文摘要:

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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