丁酸钠通过诱导铁死亡抑制肝癌细胞增殖  

SODIUM BUTYRATE INHIBITS THE PROLIFERATION OF HEPATOCELLULAR CARCINOMACELLS BY INDUCING FERROPTOSIS

作　　者：孙小蝶 秦勇 刘璐琳 张秋玉 卞中博 孙素霞[1] SUN Xiao-die;QIN Yong;LIU Lu-lin;ZHANG Qiu-yu;BIAN Zhong-bo;SUN Su-xia(Department of Nutrition and Food Hygiene,School of Public Health,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学公共卫生学院营养与食品卫生学系,广州510515

出　　处：《营养学报》2023年第2期157-162,共6页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.81773429);广东省自然科学基金(No.2214050005471)。

摘　　要：目的 探讨丁酸钠(sodium butyrate,NaB)抑制肝癌细胞增殖并诱导铁死亡的作用及其分子机制。方法分别用MTT、平板克隆和细胞形态观察探讨NaB对肝癌HepG2细胞增殖的影响。总活性氧(ROS)荧光探针DCFH-DA、脂质ROS荧光探针C11-BODIPY581/591、还原型谷胱甘肽(GSH)和MTT探讨NaB是否诱导肝癌HepG2细胞铁死亡。通过分析公共数据库TCGA、HCCDB探讨肝癌和正常组织SLC7A11表达水平以及肝癌人群的生存曲线。Westernblot检测探讨NaB对HepG2细胞SLC7A11表达的影响。结果 MTT和平板克隆实验结果显示,2mmol/LNaB显著抑制HepG2细胞增殖活性(P<0.05)和克隆球形成(P<0.05)且NaB可使细胞皱缩变圆和细胞数量显著减少。ROS和GSH检测结果显示,NaB可明显增加HepG2细胞总ROS和脂质ROS含量并下调GSH,铁死亡抑制剂(ferrostatin-1,Fer-1)可逆转NaB诱导的HepG2细胞ROS积累、GSH减少和增殖活性抑制。TCGA和HCCDB数据库分析结果显示,与正常组织相比,肝癌组织中SLC7A11显著上调(P<0.001),且SLC7A11高表达比低表达的肝癌患者预后生存期短。Western blot结果显示,NaB可下调HepG2细胞SLC7A11蛋白表达水平,Fer-1可逆转NaB诱导的SLC7A11蛋白水平下调(P<0.05)。结论 NaB通过诱导铁死亡抑制肝癌细胞增殖。[营养学报,2023,45(2):157-162]Objective To investigate the mechanism by which sodium butyrate(NaB)inhibits the proliferation of hepatocellular carcinoma cells(HCC)via inducing ferroptosis.Methods MTT assay,plate cloning,inverted light microscope were used to investigate the effects of NaB on HepG2 cells viability.The DCFH-DA,C11-BODIPY581/591 fluorescent probe,GSH,MTT assay were used to explore the ferroptosis induced by NaB in HepG2 cells.The expression level of SLC7A11 in hepatocellular carcinoma tissues and survival curve of hepatocellular carcinoma patients from TCGA and HCCDB databases were analyzed.Western blot was used to investigate the effect of NaB on the expression of SLC7A11 in HepG2 cells.Results The MTT and plate cloning tests showed that 2 mmol/L NaB significantly inhibited the proliferation of HepG2 cells(P<0.05)and the formation of clone spheres(P<0.05).NaB could significantly shrink the cells and reduce the number of cells.NaB significantly increased the total ROS and lipid ROS contents of HepG2 cells.Ferroptosis inhibitors(Ferrostatin-1,Fer-1)reversed NaB-induced inhibition on the proliferation of HepG2 cells,ROS accumulation and GSH decrease.The results of TCGA and HCCDB databases analysis showed that SLC7A11 was highly expressed in hepatocellular carcinoma tissues(P<0.001),and the survival time of patients with high expression of SLC7A11 was shorter than that of patients with low expression of hepatocellular carcinoma.Western blot test showed that NaB could down-regulate the protein level of SLC7A11 in HepG2 cells,and Fer-1 could reverse the down-regulation of SLC7A11 protein level induced by NaB(P<0.05).Conclusion It is suggested that NaB inhibits the proliferation of hepatocellular carcinoma cells by inducing ferroptosis.[ACTA NUTRIMENTA SINICA,2023,45(2):157-162]

关 键 词：丁酸钠 肝癌 铁死亡 

分 类 号：R151.2[医药卫生—营养与食品卫生学]