Wang Can, Wang Junyang, Xing Fei, Liu Ning, Wang Xudong
Department of Gastrointestinal Surgery, The Second Hospital of Jilin University, Changchun, Jilin, China.
Central Lab, The Second Hospital of Jilin University, Changchun, Jilin, China.
Front Oncol. 2025 Jun 12;15:1558290. doi: 10.3389/fonc.2025.1558290. eCollection 2025.
5-Fluorouracil (5-FU) resistance is considered to be a possible reason for the failure of conventional drug treatment of colorectal cancer (CRC). Recently, salinomycin (SAL), as a selective inhibitor of cancer stem cells (CSCs), has been used to sensitize and attenuate a variety of solid tumor chemotherapy drugs. In our study, our goal was to combine SAL with 5-FU to explore not only whether there is an increase in sensitivity of CRC to 5-FU but also the molecular mechanism involved in enhancing 5-FU sensitivity and promoting tumor cell chemotherapeutic death.
ComboSyn software was used to study whether dual drug combinations synergistically promote each other and their dosage. CCK8, EdU, and Annexin V/PI assays were used to study the cell proliferation and apoptosis of SW480 and HCT116 cells in response to SAL single-drug and dual-drug co-treatment. Cell cycle staining was used to assess cycle arrest. Wound healing and migration and invasion experiments were used to identify changes in migration and invasion capabilities under the influence of different drugs. Transcriptome sequencing is used to explore the molecular mechanisms of drugs. Reactive oxygen species (ROS) fluorescence staining and malondialdehyde (MDA) level measurement were used to confirm the changes in ferroptosis levels of SW480 and HCT116 cells after drug treatment. Nude xenograft mice were used to detect antitumor . Changes in the protein level expression of ferroptosis GPX4 and SLC7A11 were also determined in the treated cells.
SAL alone and in combination with 5-FU were found to significantly increase cell mortality and apoptosis. At the same time, our results show that the combination of SAL and 5-FU not only inhibits the proliferation, migration, and invasion of CRC cell lines and , but also promotes ferroptosis of CRC cell lines by downregulating the expression of GPX4 and SLC7A11. It may provide more and novel solutions and treatment perspectives for 5-FU or other drug-resistant chemotherapy strategies for patients with CRC.
SAL inhibits CRC, whose effect is achieved by reducing GPX4 and SLC7A11 protein levels to mediate ferroptosis activation in collaboration with 5-FU.
5-氟尿嘧啶（5-FU）耐药被认为是结直肠癌（CRC）传统药物治疗失败的一个可能原因。最近，沙林霉素（SAL）作为一种癌症干细胞（CSC）的选择性抑制剂，已被用于使多种实体瘤化疗药物增敏并减弱其毒性。在我们的研究中，我们的目标是将SAL与5-FU联合使用，不仅探索CRC对5-FU的敏感性是否增加，而且探索增强5-FU敏感性和促进肿瘤细胞化疗死亡所涉及的分子机制。
使用ComboSyn软件研究双药组合是否协同促进彼此作用及其剂量。使用CCK8、EdU和Annexin V/PI检测法研究SW480和HCT116细胞在SAL单药和双药联合处理下的细胞增殖和凋亡情况。使用细胞周期染色评估细胞周期阻滞。使用伤口愈合以及迁移和侵袭实验来确定不同药物影响下迁移和侵袭能力的变化。使用转录组测序探索药物的分子机制。使用活性氧（ROS）荧光染色和丙二醛（MDA）水平测量来确认药物处理后SW480和HCT116细胞铁死亡水平的变化。使用裸鼠异种移植模型检测抗肿瘤作用。还测定了处理后细胞中铁死亡相关蛋白GPX4和SLC7A11的水平变化。
发现单独使用SAL以及SAL与5-FU联合使用均能显著增加细胞死亡率和凋亡。同时，我们的结果表明，SAL与5-FU联合使用不仅抑制CRC细胞系SW480和HCT116的增殖、迁移和侵袭，还通过下调GPX4和SLC7A11的表达促进CRC细胞系的铁死亡。这可能为CRC患者的5-FU或其他耐药化疗策略提供更多新颖的解决方案和治疗视角。
SAL抑制CRC，其作用是通过降低GPX4和SLC7A11蛋白水平，与5-FU协同介导铁死亡激活来实现的。
