PolyphyllinⅥ诱导铁死亡抑制结肠癌细胞株HT-29和SW480增殖机制研究  

Study on the mechanism of polyphyllin Ⅵ in inhibiting proliferation of HT-29 and SW480 colon cancer cell lines by inducing ferroptosis

作　　者：李斌[1] 易小江 Li Bin;Yi Xiaojiang(Department of General Surgery,Shenzhen Hospital,Bejing University of Chinese Medicine,Shenzhen 518172,Guangdong,China;Department of Colorectal Surgery,Gastrointestinal Tumor Center,Second Affliated Hospital of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Traditional Chinese Medicine),Guangzhou 510120,Guangdong,China)

机构地区：[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

出　　处：《消化肿瘤杂志(电子版)》2025年第1期56-64,共9页Journal of Digestive Oncology(Electronic Version)

基　　金：广东省中医药管理局科研项目(20231148)。

摘　　要：目的探讨重楼皂苷polyphyllinⅥ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、polyphyllinⅥ组(用浓度为5μmol/LpolyphyllinⅥ处理的细胞)和polyphyllinⅥ+ferrostatin-1组(用浓度为5μmol/LpolyphyllinⅥ+100μumol/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polyphylin Ⅵ组中的表达水平。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polyphylinⅥ可抑制细胞株HT-29和SW480的增殖活力。与阴性对照组相比,polyphyllinⅥ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polyphyllinⅥ组细胞株的线粒体数量减少,膜密度增高。与polyphyllin Ⅵ组相比,polyphyllinⅥ+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 Asynthase 4,ACSL4)蛋白和mRNA表达水平在polyphyllinⅥ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PolyphyllinⅥ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。Objective To investigatethemechanismof polyphyllin Ⅵ in inhibitingtheproliferation 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 V group(cells treated with a concentration of 5μmol/L polyphylin Ⅵ),and polyphyllin Ⅵ+ferrostatin-1 group(cells treated with a concentration of 5μmol/L polyphyllin Ⅵ+100μmol/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 Ⅵgroup 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 feroptosis 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

关 键 词：PolyphylinⅥ 结肠癌 细胞株HT-29和SW480 铁死亡 

分 类 号：R73[医药卫生—肿瘤]