国际口腔医学杂志

国际口腔医学杂志 ›› 2026, Vol. 53 ›› Issue (3): 352-361.doi: 10.7518/gjkq.2026213

• 论著 • 上一篇    

葫芦素B通过核因子E2相关因子2/溶质载体家族7成员11/谷胱甘肽过氧化物酶4信号通路诱导舌鳞状细胞癌CAL-27细胞铁死亡

林健辉(),蒋文芮,韩瑞,刘新伟,张靓,周美云,徐锦程()   

  1. 蚌埠医科大学第一附属医院口腔科 蚌埠 233004
  • 收稿日期:2024-11-01 修回日期:2025-10-20 出版日期:2026-05-01 发布日期:2026-04-24
  • 通讯作者: 徐锦程
  • 作者简介:林健辉，硕士，Email：839606085@qq.com
  • 基金资助:
    安徽省高校自然科学研究项目(KJ2021ZD0088)

Cucurbitacin B induces ferroptosis in CAL-27 cells derived from tongue squamous cell carcinoma through the signaling pathway involving nuclear factor erythroid 2-related factor 2, solute carrier family 7 member 11, and glutathione peroxidase 4

Jianhui Lin(),Wenrui Jiang,Rui Han,Xinwei Liu,Liang Zhang,Meiyun Zhou,Jincheng Xu()   

  1. Dept. of Stomatology, the First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
  • Received:2024-11-01 Revised:2025-10-20 Online:2026-05-01 Published:2026-04-24
  • Contact: Jincheng Xu
  • Supported by:
    Natural Science Research Projects of Colleges and Universities in Anhui Province(KJ2021ZD0088)

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摘要：

目的 观察葫芦素B（CuB）是否诱导舌鳞状细胞癌CAL-27细胞铁死亡，并探讨其可能的机制。 方法 采用不同浓度的CuB（0、5、10、15、20、30 µmol/L）处理CAL-27细胞，使用细胞计数试剂盒（CCK）-8法检测细胞增殖活力，并计算半数抑制浓度（IC50）；细胞克隆实验、细胞划痕实验用于检测不同浓度CuB对CAL-27细胞增殖、迁移的影响；采用不同浓度CuB处理CAL-27细胞，分别检测细胞内活性氧（ROS）、Fe2+、谷胱甘肽（GSH）、丙二醛（MDA）的含量变化；蛋白印迹法（WB）检测核因子E2相关因子2（Nrf2）、溶质载体家族7成员11（SLC7A11）及谷胱甘肽过氧化物酶4（GPX4）蛋白的表达水平。 结果 CCK-8结果显示：5、10、15、20、30 µmol/L的CuB能显著抑制CAL-27细胞增殖活力，IC50为13.93 µmol/L（P<0.001）。平板克隆实验显示：CuB能够抑制CAL-27细胞的克隆形成能力（P<0.01）。划痕实验显示：葫芦素显著缩短CAL-27细胞的迁移距离抑制迁移能力（P<0.01）。ROS、GSH、MDA及Fe2+测定结果显示：与对照组相比，不同浓度CuB干预后CAL-27细胞中ROS、Fe2+的荧光强度显著增强，且细胞内GSH含量明显下降，MDA含量则明显上升，诱导细胞氧化损伤，而以上变化可被铁死亡抑制剂铁抑素-1（Ferrostatin-1）逆转（P<0.01）。WB结果显示：不同浓度CuB干预后能够显著下调Nrf2、SLC7A11和GPX4蛋白表达水平（P<0.05），且铁死亡抑制剂Fer-1可逆转Nrf2、SLC7A11、GPX4蛋白表达（P<0.05）。 结论 CuB可诱导舌鳞状细胞癌CAL-27细胞发生铁死亡，其机制可能与CuB通过调节Nrf2/SLC7A11/GPX4信号通路有关。

关键词: 葫芦素B, 铁死亡, 舌鳞状细胞癌, 核因子E2相关因子2/溶质载体家族7成员11/谷胱甘肽过氧化物酶4信号通路

Abstract:

Objective This study aims to observe whether Cucurbitacin B (CuB) induces ferroptosis in CAL-27 cells derived from tongue squamous cell carcinoma and explore its possible mechanism. Methods CAL-27 cells were treated with different concentrations of CuB (0, 5, 10, 15, 20, and 30 μmol/L). Cell proliferation activity was detected using the cell counting kit-8 (CCK-8) method, and the median inhibition concentration (IC50) was calculated. Cell clo-ning and wound healing assays were used to detect the effects of different concentrations of CuB on the proliferation and migration of CAL-27 cells. CAL-27 cells were treated with different concentrations of CuB, and the changes in intracellular reactive oxygen species (ROS), Fe2+, GSH, and malondialdehyde (MDA) levels were detected. Western blot (WB) was used to detect the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) proteins. Results CCK-8 results showed that CuB at concentrations of 5, 10, 15, 20, and 30 μmol/L significantly inhibited the proliferation activity of CAL-27 cells, with an IC50 of 13.93 μmol/L (P<0.001). The plate cloning assay demonstrated that CuB inhibited the cloning ability of CAL-27 cells (P<0.01). The wound healing assay revealed that CuB significantly shortened the migration distance of CAL-27 cells and inhibited their migration ability (P<0.01). The results of ROS, GSH, MDA, and Fe2+ measurements indicated that, compared with the control group, the fluorescence intensity of ROS and Fe2+ in CAL-27 cells significantly increased after CuB intervention at different concentrations. Meanwhile, the intracellular GSH content significantly decreased, and the MDA content significantly increased, which induced cellular oxidative damage. These changes could be reversed by the ferroptosis inhibitor Fer-1 (P<0.01). WB results showed that CuB intervention at different concentrations significantly downregulated the expression levels of Nrf2, SLC7A11, and GPX4 proteins (P<0.05). Furthermore, the ferroptosis inhibitor Fer-1 reversed the protein expression of Nrf2, SLC7A11 and GPX4 (P<0.05). Conclusion CuB can induce ferroptosis in CAL-27 cells derived from tongue squamous cell carcinoma, and its mechanism may be related to the ability of CuB to regulate the signaling pathway involving Nrf2, SLC7A11, and GPX4.

Key words: cucurbitacin B, ferroptosis, oral squamous cell carcinoma, nuclear factor erythroid 2-related factor 2/solute carrier family 7 member 11/glutathione peroxidase 4 signaling pathway

中图分类号: 

  • R782

图 1

不同浓度CuB对CAL-27细胞增殖活性的影响与对照组相比，***P<0.001。"

图 2

不同浓度CuB对CAL-27细胞迁移能力的影响A：各组细胞平板划痕图（倒置显微镜 × 100）；B：各组细胞平板划痕统计图与对照组相比，**P<0.01，***P<0.001。"

图 3

不同浓度CuB对CAL-27细胞克隆能力的影响A：各组细胞平板克隆图；B：各组细胞平板克隆统计图，与对照组相比，**P<0.01，***P<0.001。"

图 4

CuB对CAL-27细胞铁死亡相关指标的影响A：各组细胞ROS荧光强度图 荧光显微镜 × 200；B：各组细胞Fe2+荧光强度图 荧光显微镜 × 200；C：各组细胞ROS荧光强度统计图；D：各组细胞Fe2+荧光强度统计图；E：各组细胞GSH含量统计图；F：各组细胞MDA含量统计图；与对照组相比，**P<0.01，***P<0.001。"

图 5

Fer-1联合CuB对CAL-27细胞铁死亡相关指标的影响A：各组细胞ROS荧光强度图 荧光显微镜 × 200；B：各组细胞Fe2+荧光强度图 荧光显微镜 × 200；C：各组细胞ROS荧光强度统计图；D：各组细胞Fe2+荧光强度统计图；E：各组细胞GSH含量统计图；F：各组细胞MDA含量统计图；与对照组相比，**P<0.01，***P<0.001。"

图 6

CuB对CAL-27细胞铁死亡相关蛋白表达的影响A：各组细胞中铁死亡相关分子蛋白表达图；B~D：各组细胞中Nrf2、SLC7A11、GPX4的蛋白表达统计图；与对照组相比，*P<0.05，**P<0.01，***P<0.001。"

图 7

Fer-1联合CuB对CAL-27细胞铁死亡相关蛋白表达的影响A：各组细胞中铁死亡相关分子蛋白表达图；B~D：各组细胞中Nrf2、SLC7A11、GPX4的蛋白表达统计图；与对照组相比，*P<0.05，**P<0.01，***P<0.001。"

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