硒代胱氨酸通过诱导活性氧产生启动氧化应激损伤抑制结肠癌细胞生长

doi:10.12122/j.issn.1673-4254.2026.03.07

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (3): 532-540.doi: 10.12122/j.issn.1673-4254.2026.03.07

• 基础研究 • 上一篇

硒代胱氨酸通过诱导活性氧产生启动氧化应激损伤抑制结肠癌细胞生长

宋淇乐¹(), 苗益恺², 冯小桐¹, 王一凡¹, 刘伟¹, 魏琪¹, 于新汝¹, 陈文文³(), 付晓艳⁴()

^1.山东省医药卫生脑损伤与功能康复重点实验室//山东第一医科大学第二附属医院神经内科，山东泰安 271000
^2.山东省临沂市人民医院研究生培养基地//广州中医药大学，广东广州 510000，山东第一医科大学3. 第二附属医院药学部
^4.临床与基础医学院，山东泰安 271000

收稿日期:2025-09-23 出版日期:2026-03-20 发布日期:2026-03-26
通讯作者: 陈文文,付晓艳 E-mail:sqlbangong@163.com;wen-860521@163.com;txyfu66@163.com
作者简介:宋淇乐，在读硕士研究生，E-mail: sqlbangong@163.com
基金资助:
国家自然科学基金(81701178);山东省医药卫生科技项目(202513011302)

Selenocystine inhibits colon cancer cell growth by promoting reactive oxygen species generation to trigger oxidative damage

Qile SONG¹(), Yikai MIAO², Xiaotong FENG¹, Yifan WANG¹, Wei LIU¹, Qi WEI¹, Xinru YU¹, Wenwen CHEN³(), Xiaoyan FU⁴()

^1.Department of Neurology, Shandong Provincial Key Laboratory of Brain Injury and Functional Rehabilitation, Second Affiliated Hospital of Shandong First Medical University, Taian 271000, China
^2.Postgraduate Training Base of Linyi People's Hospital of Shandong Province, Guangzhou University of Chinese Medicine, Guangzhou 510000, China
^3.Department of Pharmacy, Second Affiliated Hospital
^4.School of Clinic and Basic Medicine, Shandong First Medical University, Taian 271000, China

Received:2025-09-23 Online:2026-03-20 Published:2026-03-26
Contact: Wenwen CHEN, Xiaoyan FU E-mail:sqlbangong@163.com;wen-860521@163.com;txyfu66@163.com
Supported by:
National Natural Science Foundation of China(81701178)

摘要/Abstract

摘要：

目的评价硒代胱氨酸（SeC）体外抑制结肠癌细胞生长的分子机制。方法体外培养RKO、HCT-116、LoVo结肠癌细胞，SeC处理细胞24 h和48 h，分为正常对照组和SeC低、中、高剂量组（SeC为5、10、20 μmol/L）。MTT检测结肠癌细胞的活性。划痕实验检测结肠癌细胞迁移能力。流式细胞仪PI染色检测细胞周期阻滞和凋亡情况。荧光探针检测自由基产生、线粒体结构和膜电位变化。铁死亡标志物如丙二醛（MDA）、谷胱甘肽（GSH）、亚铁离子（Fe²⁺）分析铁死亡的水平，Western blotting检测蛋白表达情况。结果 SeC低、中、高剂量组均抑制结肠癌细胞增殖和迁移（P<0.05）；SeC不同剂量处理组中细胞周期相关蛋白CDK2、CDK4表达下调，并且凋亡蛋白PARP和Caspase9发生活化（P<0.05）；此外，Western blotting结果显示SeC诱导铁死亡蛋白FTH1，xCT表达下降和DMT1表达升高；SeC不同剂量处理组中MDA，Fe²⁺水平增加和GSH水平降低（P<0.05）。荧光结果显示线粒体结构损伤、活性氧（ROS）生成增多；Western blotting结果显示氧化损伤蛋白的磷酸化表达升高和NRF2/HO-1蛋白表达下调（P<0.05）；ROS清除剂显著逆转结肠癌细胞中SeC对DMT1、PARP、p-H₂A.X蛋白的上调作用（P<0.05）。结论 SeC可通过诱导ROS生成启动氧化应激损伤诱导结肠癌细胞凋亡和铁死亡，进而抑制结肠癌细胞生长，是结肠癌潜在的化疗剂。

关键词: 硒代胱氨酸, 结肠癌, 活性氧, 氧化应激, 凋亡, 铁死亡

Abstract:

Objective To explore the molecular mechanism by which selenocystine (SeC) inhibits colon cancer cell growth in vitro. Methods Colon cancer cells (RKO, HCT-116, and LoVo) were cultured and treated with 5, 10, or 20 μmol/L SeC for 24 h and 48 h. MTT assay was used to detect the cell viability, and wound healing assay was used to examine changes in cell migration. Flow cytometry with PI staining was used to analyze cell cycle arrest and apoptosis. Fluorescence probes were employed to monitor reactive oxygen species (ROS) generation, mitochondrial morphology and membrane potential, and the changes in ferroptosis were evaluated by detecting malondialdehyde (MDA), glutathione (GSH) and ferrous ion (Fe²⁺) levels; Western blotting was used to detect the changes in protein expressions. Results SeC at all the 3 doses significantly inhibited proliferation and migration of colon cancer cells, down-regulated the expression of cell cycle-related proteins CDK2 and CDK4 and activated the apoptotic proteins PARP and caspase-9. Western blotting showed that SeC decreased the expression of ferroptosis proteins FTH1 and xCT and increased the expression of DMT1. The levels of MDA and Fe²⁺ were increased and GSH level was decreased in SeC-treated cells. Fluorescence staining results showed that SeC treatment induced mitochondrial structure damages and promoted cellular ROS production. SeC treatment also increased phosphorylation of oxidative damage proteins and lowered the expression levels of NRF2 and HO-1 proteins. ROS scavenger significantly reversed the up-regulation of DMT1, PARP and p-H₂A.X protein induced by SeC in colon cancer cells. Conclusion SeC induces apoptosis and ferroptosis of colon cancer cells by promoting ROS generation and initiating oxidative damage, suggesting the potential of SeC as a potential chemotherapeutic agent for colon cancer.

Key words: selenocystine, colon cancer, reactive oxygen species, oxidative stress, apoptosis, ferroptosis

宋淇乐, 苗益恺, 冯小桐, 王一凡, 刘伟, 魏琪, 于新汝, 陈文文, 付晓艳. 硒代胱氨酸通过诱导活性氧产生启动氧化应激损伤抑制结肠癌细胞生长[J]. 南方医科大学学报, 2026, 46(3): 532-540.

Qile SONG, Yikai MIAO, Xiaotong FENG, Yifan WANG, Wei LIU, Qi WEI, Xinru YU, Wenwen CHEN, Xiaoyan FU. Selenocystine inhibits colon cancer cell growth by promoting reactive oxygen species generation to trigger oxidative damage[J]. Journal of Southern Medical University, 2026, 46(3): 532-540.

图/表 5

图1 SeC剂量依赖性抑制结肠癌细胞的生长

Fig.1 SeC inhibits growth of colon cancer cells in a dose-dependent manner. A: SeC inhibits viability of RKO, HCT-116 and LoVo cells detected by MTT assay. B: Changes in cell morphology (Original magnification: ×200). C: SeC inhibits the migration of RKO cells detected by wound healing assay. (×100). D: Quantitative analysis of cell migration. All experiments were repeated 3 times (n=3). All data were presented as Mean±SD. *P<0.05, **P<0.01 vs SeC 0 μmol/L.

图2 SeC诱导结肠癌细胞周期阻滞和凋亡

Fig.2 SeC induces cell cycle arrest and apoptosis in colon cancer cells. A: SeC inhibits expressions of RKO cell cycle-related proteins detected by Western blotting. B: SeC-induced cell cycle arrest and apoptosis in RKO, HCT-116, and LoVo cells detected by flow cytometry with PI staining. C: Quantitative analysis of cell cycle proteins. D: Quantitative analysis of cell cycle arrest in RKO, HCT-116, and LoVo cells. All experiments were repeated three times (n=3). All data are presented as Mean±SD. *P<0.05 vs SeC 0 μmol/L.

图3 SeC诱导结肠癌细胞凋亡和铁死亡

Fig.3 SeC induces apoptosis and ferroptosis in colon cancer cells. A: PARP cleavage and caspase-9 activation. B: Effect of SeC on ferroptosis-related proteins in RKO cells detected by Western blotting. C: Quantitative analysis of apoptotic and ferroptosis-related proteins. D: Measurement of MDA, GSH and Fe2+ in RKO cells after SeC treatment. All experiments were repeated three times (n=3). All data are presented as Mean±SD. *P<0.05 vs SeC 0 μmol/L.

图4 SeC诱导结肠癌细胞线粒体功能障碍和氧化应激损伤

Fig.4 SeC induces mitochondrial dysfunction and oxidative stress damage in colon cancer cells. A: Effects of SeC on oxidative stress-related proteins in RKO cells detected by Western blotting. B: SeC induces ROS accumulation and mitochondrial dysfunction in RKO cells. DCFH-DA fluorescent probe was used to detect ROS production (×100). Mito-SOX fluorescent probe was used to detect mitochondrial superoxide production (×400). Mito-Tracker fluorescent probe was used to detect mitochondrial structural damage (×400). JC-1 fluorescent probe was used to detect mitochondrial membrane potential changes (×200). C: Quantitative analysis of oxidative stress-related protein and DNA damage-related protein results. D: Quantitative analysis of ROS, Mito-SOX and JC-1 fluorescence results. All experiments were repeated 3 times (n=3). All data are presented as Mean±SD. *P<0.05 vs SeC 0 μmol/L.

图5 ROS清除减弱SeC诱导的凋亡和铁死亡

Fig.5 ROS elimination attenuates SeC-induced apoptosis and ferroptosis. A: NAC pretreatment inhibits SeC-induced DNA damage, apoptosis, and ferroptosis detected by Western blotting. B: NAC pretreatment reverses the effects of SeC on colon cancer cells. C: Changes in cell morphology (×200). D: Quantitative analysis of apoptosis-related proteins, ferroptosis-related proteins, and DNA damage-related proteins. All experiments were repeated 3 times (n=3). All data are presented as Mean±SD. *P<0.05 vs SeC 0 μmol/L.

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硒代胱氨酸通过诱导活性氧产生启动氧化应激损伤抑制结肠癌细胞生长

Selenocystine inhibits colon cancer cell growth by promoting reactive oxygen species generation to trigger oxidative damage

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