| 刘笑笑,王玉琛,耿尚怡,等.瘦素通过抑制铁死亡途径介导三阴性乳腺癌放射抵抗的机制研究[J].中华放射医学与防护杂志,2026,46(1):20-28.Liu Xiaoxiao,Wang Yuchen,Geng Shangyi,et al.Mechanism of leptin in mediating radioresistance in triple-negative breast cancer through the inhibition of ferroptosis pathway[J].Chin J Radiol Med Prot,2026,46(1):20-28 |
| 瘦素通过抑制铁死亡途径介导三阴性乳腺癌放射抵抗的机制研究 |
| Mechanism of leptin in mediating radioresistance in triple-negative breast cancer through the inhibition of ferroptosis pathway |
| 投稿时间：2025-06-20 |
| DOI：10.3760/cma.j.cn112271-20250620-00207 |
| 中文关键词: 三阴性乳腺癌 瘦素 铁死亡 脂质运载蛋白2 放疗抵抗 |
| 英文关键词:Triple-negative breast cancer Leptin Ferroptosis Lipocalin 2 Radioresistance |
| 基金项目:国家自然科学基金(82172839,82374229,82573452,82574765);陕西省青年科技新星项目(2024ZC-KJXX-083) |
| 作者 | 单位 | E-mail | | 刘笑笑 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 王玉琛 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 耿尚怡 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 党永泽 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 李霞 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 郭伟 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 柯悦 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 潘书沛 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 曲敬琨 | 西安交通大学第二附属医院乳腺疾病诊疗中心, 西安 710004 | xixizhao@xjtu.edu.cn | | 马红兵 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | | | 赵茜茜 | 西安交通大学第二附属医院肿瘤放疗科, 西安 710004 | |
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| 中文摘要: |
| 目的 探讨瘦素通过调控铁死亡相关信号通路对三阴性乳腺癌放疗抵抗的影响。方法 采用不同浓度瘦素(0、50、100、200 ng/ml)处理MDA-MB-231细胞,通过蛋白质免疫印迹和细胞增殖及毒性检测方法优化处理条件。将MDA-MB-231细胞设为4组:对照组、瘦素组(200 ng/ml,48 h)、照射组(4 Gy X射线)及照射+瘦素组(联合处理组),使用酶标仪检测丙二醛(MDA)水平,流式细胞仪检测活性氧(ROS)和细胞凋亡水平。Western blot实验检测磷酸化的信号传导及转录激活蛋白3(p-STAT3)、溶质载体家族7成员11基因(SLC7A11)和谷胱甘肽过氧化物酶4(GPX4)的表达,同时结合生物信息学(GEO和FerrDb数据库)筛选关键分子并验证脂质运载蛋白2(LCN2)的表达。通过 siRNA 敲低 LCN2,评估其对瘦素调控 SLC7A11/GPX4 的作用,同时在乳腺癌组织中进行免疫组织化学分析。结果 与0 ng/ml相比,200 ng/ml瘦素作用48 h后可显著增加 MDA-MB-231 细胞的p-STAT3的表达水平和细胞增殖(t = 32.85、17.25,P < 0.05)。与对照组相比,瘦素显著降低了MDA-MB-231 细胞的凋亡率(t = -6.87,P < 0.05),并且明显减弱了照射诱导的细胞凋亡(t = -9.46,P < 0.05)。与照射组相比,瘦素可抑制照射诱导MDA-MB-231细胞的MDA 和 ROS 水平(t= -4.67、 -14.82,P < 0.05),同时上调铁死亡抑制蛋白SLC7A11和GPX4的表达(t = 7.11、8.36,P < 0.05),从而部分逆转照射诱导的铁死亡。在照射条件下,瘦素显著上调MDA-MB-231细胞的 LCN2 表达(t = 8.96,P < 0.05),而 LCN2 被 siRNA 敲低后,SLC7A11 与 GPX4 的上调效应被逆转(t = -5.85、-15.58,P < 0.05),提示瘦素通过 LCN2 维持 SLC7A11/GPX4 的稳定性,从而抑制铁死亡并增强放疗抵抗。免疫组织化学结果显示,高瘦素表达的乳腺癌组织中,LCN2、SLC7A11 与 GPX4 水平均显著升高(t = 50.57、10.14、13.81,P < 0.05)。结论 瘦素可通过LCN2/SLC7A11/GPX4信号通路抑制铁死亡,参与三阴性乳腺癌放疗抵抗,为靶向铁死亡的放射增敏策略提供潜在依据。 |
| 英文摘要: |
| Objective To investigate the effect of leptin on radiotherapy resistance in triple-negative breast cancer by regulating ferroptosis-related signaling pathways. Methods Different concentrations of leptin (0, 50, 100, 200 ng/ml) were used to treat MDA-MB-231 cells, and the treatment conditions were optimized through western blot analysis and cell proliferation and toxicity assays. MDA-MB-231 cells were divided into four groups: control group, leptin group (200 ng/ml, 48 h), irradiation group (4 Gy X-rays), and irradiation + leptin group. Malondialdehyde (MDA) levels were detected using a microplate reader, and reactive oxygen species (ROS) and cell apoptosis levels were measured using flow cytometry. Western blotting was used to detect the expression of phosphorylated signal transducer and activator of transcription 3 (p-STAT3), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4). Key molecules were screened using bioinformatics (GEO and FerrDb databases) and the expression of lipid carrier protein 2 (LCN2) was validated. siRNA was used to knock down LCN2, and its effect on leptin-regulated SLC7A11/GPX4 expression was evaluated. Immunohistochemistry was performed on breast cancer tissues. Results Compared to the 0 ng/ml, 48 h group, leptin (200 ng/ml, 48 h) significantly increased the expression level of p-STAT3 and cell proliferation in MDA-MB-231 cells (t = 32.85, 17.25, P < 0.05). Compared to the control group, leptin significantly reduced the apoptosis rate of MDA-MB-231 cells (t = -6.87, P < 0.05), and markedly attenuated irradiation-induced apoptosis (t = -9.46, P < 0.05). Compared to the irradiation group, leptin inhibited the MDA and ROS levels induced by radiation in MDA-MB-231 cells (t = -4.67, -14.82, P < 0.05), and simultaneously upregulated the expression of ferroptosis-suppressing proteins SLC7A11 and GPX4 (t = 7.11, 8.36, P < 0.05), partially reversing radiation-induced ferroptosis. Mechanistic studies showed that under irradiation conditions, leptin significantly upregulated the expression of LCN2 in MDA-MB-231 cells (t = 8.96, P < 0.05). After LCN2 was knocked down by siRNA, the upregulation effects of SLC7A11 and GPX4 were reversed (t = -5.85, -15.58, P < 0.05), suggesting that leptin maintains the stability of SLC7A11/GPX4 through LCN2, thereby inhibiting ferroptosis and enhancing radiotherapy resistance. Immunohistochemical analysis further showed that in breast cancer tissues with high leptin expression, the levels of LCN2, SLC7A11, and GPX4 were significantly elevated (t = 50.57, 10.14, 13.81, P < 0.05). Conclusion Leptin inhibits ferroptosis through the LCN2/SLC7A11/GPX4 signaling pathway, contributing to radioresistance in triple-negative breast cancer, and may serve as a potential target for ferroptosis-based radiosensitization strategies. |
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