引用文本：李斌, 易小江. Polyphyllin Ⅵ诱导铁死亡抑制结肠癌细胞株HT-29和SW480增殖机制研究[J/CD]. 消化肿瘤杂志（电子版）, 2025, 17(1):56-64.

作者：李斌1，易小江2

单位：1. 北京中医药大学深圳医院 普通外科，广东 深圳 518172；2. 广州中医药大学第二附属医院（广东省中医院） 胃肠肿瘤中心结直肠外科，广东 广州 510120

Authors：Li Bin1, Yi Xiaojiang2

Unit：1. Department of General Surgery, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen 518172, Guangdong, China；2. Department of Colorectal Surgery, Gastrointestinal Tumor Center, Second Affiliated Hospital of Guangzhou University of Chinese Medicine (Guangdong Provincial Hospital of Traditional Chinese Medicine), Guangzhou 510120, Guangdong, China

摘要：

目的 探讨重楼皂苷polyphyllin Ⅵ通过诱导铁死亡抑制结肠癌细胞株HT-29和SW480增殖的相关机制。方法 常规培养细胞株HT-29和SW480，实验分为阴性对照组、二甲基亚砜组、polyphyllin Ⅵ组（用浓度为5 μmol/L polyphyllin Ⅵ处理的细胞）和polyphyllin Ⅵ+ferrostatin-1组（用浓度为5 μmol/L polyphyllin Ⅵ+100 μmol/L铁死亡抑制剂ferrostatin-1联合处理的细胞）。采用高通量转录组测序分析polyphyllin Ⅵ组的差异表达基因并进行基因富集分析。采用CCK-8法检测各组细胞株的增殖活力。采用活性氧、铁浓度、谷胱甘肽浓度检测实验和透射电镜检测分析各组细胞株的铁死亡改变。最后采用蛋白质印迹法和定量聚合酶链反应分别验证铁死亡相关蛋白和基因在polyphyllin Ⅵ组的表达水平。结果 转录组测序和基因富集分析显示polyphyllin Ⅵ可通过铁死亡通路作用于细胞株HT-29和SW480。CCK-8法结果显示polyphyllin Ⅵ可抑制细胞株HT-29和SW480的增殖活力。与阴性对照组相比，polyphyllin Ⅵ组的活性氧水平增加（P＜0.001）、谷胱甘肽浓度降低（P＜0.05）、铁离子相对浓度增加（P＜0.001），且透射电镜检测显示polyphyllin Ⅵ组细胞株的线粒体数量减少，膜密度增高。与polyphyllin Ⅵ组相比，polyphyllin Ⅵ+ferrostatin-1组的细胞株增殖活力增强，活性氧水平降低（P＜0.001）、谷胱甘肽浓度增加（P＜0.01）、铁离子相对浓度降低（P＜0.001），且细胞株的线粒体数量上升，膜密度降低。长链酰基辅酶A合酶4（long-chain acyl-coenzyme A synthase 4, ACSL4）蛋白和mRNA表达水平在polyphyllin Ⅵ组中均上升，而谷胱甘肽过氧化物酶（glutathione peroxidases，GPX）4的蛋白表达水平则下调。结论 Polyphyllin Ⅵ可通过诱导铁死亡过程来抑制结肠癌细胞株HT-29和SW480的增殖能力，潜在作用路径为GPX4/ACSL4通路。

关键词：Polyphyllin Ⅵ；结肠癌；细胞株HT-29和SW480；铁死亡

Abstract：

Objective To investigate the mechanism of polyphyllin Ⅵ in inhibiting the proliferation of colon cancer cell lines HT-29 and SW480 by inducing ferroptosis. Method The HT-29 and SW480 cell lines were cultured routinely and divided into negative control group, dimethyl sulfoxide group, polyphyllin Ⅵ group (cells treated with a concentration of 5 μmol/L polyphyllin Ⅵ), and polyphyllin Ⅵ+ferrostatin-1 group (cells treated with a concentration of 5 μmol/L polyphyllin Ⅵ+100 μmol/L ferroptosis inhibitor ferrostatin-1). Using high-throughput transcriptome sequencing to analyze differentially expressed genes in the polyphyllin Ⅵ group and conducting gene enrichment analysis. Using CCK-8 assay to detect the proliferation activity of cell lines in each group. Reactive oxygen species, iron concentration, glutathione concentration detection experiments and transmission electron microscopy were conducted to analyze the changes in ferroptosis in each group of cell lines. Finally, western blot and quantitative polymerase chain reaction were used to verify the expression levels of ferroptosis related proteins and genes in the polyphyllin Ⅵ group, respectively. Result The results of transcriptome sequencing and gene enrichment analysis showed that polyphyllin Ⅵ can act on cell lines HT-29 and SW480 through the ferroptosis pathway. The results of CCK-8 assay showed that polyphyllin Ⅵ could inhibit the proliferation activity of cell lines HT-29 and SW480. Compared with the negative control group, the level of reactive oxygen species increased (P<0.001), the concentration of glutathione decreased (P<0.05), and the relative concentration of iron ions increased (P<0.001) in the polyphyllin Ⅵ group. Transmission electron microscopy analysis showed a decrease in mitochondrial count and an increase in membrane density in the cell lines of polyphyllin Ⅵ group. Compared with the polyphyllin Ⅵ group, the cell lines proliferation activity of the polyphyllin Ⅵ+ferrostatin-1 group was enhanced, the level of reactive oxygen species was reduced (P<0.001), the concentration of glutathione was increased (P<0.01), the relative concentration of iron ions was reduced (P<0.001), and the mitochondrial count increased while the membrane density decreased in the cell lines of polyphyllin Ⅵ+ferrostatin-1 group. The protein and mRNA expression levels of long-chain acyl-coenzyme A synthase 4 (ACSL4) increased in the polyphyllin Ⅵ group, while the protein expression level of glutathione peroxidase (GPX) 4 was downregulated. Conclusion Polyphyllin Ⅵ can inhibit the proliferation ability of the HT-29 and SW480 colon cancer cell lines by inducing ferroptosis, with a potential GPX4/ASCL4 pathway.

Key words：Polyphyllin Ⅵ; Colon cancer; HT-29 and SW480 cell lines; Ferroptosis

注：网络优先发布