摘要
目的探究尼克酰胺转甲基酶介导细胞内活性氧(reactive oxygen species,ROS)在肝癌细胞铁死亡中的影响及其机制。方法选取2019年3月至2020年2月于笔者医院接受治疗的40例原发性肝细胞癌患者作研究对象,分别取患者的癌旁组织标本及肝癌组织标本。使用串联液相质谱仪检测细胞中甲基尼克酰胺(methyl nicotinamid,MNA)表达,使用流式细胞仪检测ROS及过氧化脂质的平均荧光强度,采用蛋白质印迹法(Western blot)法检测人肝癌细胞(SK-Hep-1、Hep3B)细胞表达的情况变化。采用免疫组化法检测癌旁组织、肝癌组织中尼克酰胺转甲基酶(nicotinamide N-methyltransferase,NNMT)及ROS水平,采用CCK-8法检测不同铁浓度细胞的生存活性。结果肝癌组织组中的MNA水平较癌旁组织组有所提升差异有统计学意义(P<0.05)。与癌旁组织组比较,肝癌组织组的ROS平均荧光强度表达升高,过氧化脂质平均的荧光强度表达降低,SK-Hep-1、Hep3B细胞表达量上升,差异有统计学意义(P<0.05)。与对照组比较,NNMT组2、10、20及25μmol/L的细胞生存活性水平升高(P<0.05)。与NNMT组比较,铁抑制组不同铁浓度(2μmol/L、10μmol/L、20μmol/L、25μmol/L)的细胞生存活性表达下降差异有统计学意义(P<0.05)。结论经尼克酰胺转甲基酶介导可引导产生ROS及能量紊乱,提高了肿瘤细胞的死亡率。
Objective To explore the effect of nicotinamide transmethylase on intracellular reactive oxygen species(ROS)in iron death of hepatocellular carcinoma cells and its mechanism.Methods Methyl nicotinamide(MNA)expression in cells was detected using a tandem liquid chromatography-mass spectrometry.The average fluorescence intensity of ROS and lipid peroxidation was measured using a flow cytometer.Western blot was used to detect changes in the expression of human liver cancer cells(SK-Hep-1,Hep3B).Forty patients with primary hepatocellular carcinoma who received treatment in our hospital from March 2019 to February 2020 were selected as the study subjects,and their adjacent tissue samples and liver cancer tissue samples were collected.Immunohistochemical methods were used to detect the levels of nicotinamide N-methyltransferase(NNMT)and ROS in adjacent and liver cancer tissues.CCK-8 method was used to detect the survival activity of cells with different iron concentrations.Results The MNA levels in the liver cancer tissue group were higher than those in the adjacent tissue group(P<0.05).Compared with the adjacent tissue group,the average fluorescence intensity expression of ROS in the liver cancer tissue group increased,while the average fluorescence intensity expression of lipid peroxidation decreased(P<0.05).Compared with the adjacent tissue group,the expression levels of SK Hep-1 and Hep3B cells in the liver cancer tissue group increased(P<0.05).Compared with the control group,NNMT groups 2,10,20,and 25μmol/L The cell survival activity level increased(P<0.05);Compared with the NNMT group,the iron inhibition group had different iron concentrations2μmol/L,10μmol/L,20μmol/L,25μmol/L.The expression of cell viability decreased(P<0.05).Conclusion ROS mediated by nicotinamide methyltransferase can be guided to produce ROS and energy disorders,leading to increased tumor cell death.
作者
王锦春
戴永青
王亚清
陈珏
刘祖平
李烨佳
WANG Jinchun;DAI Yongqing;WANG Yaqing;CHEN Jue;LIU Zuping;LI Yejia(Department of Blood Transfusion,Affiliated Hospital of Shaoxing University,Shaoxing 312000,Zhejiang,China;Department of Pathology,Affiliated Hospital of Shaoxing University,Shaoxing 312000,Zhejiang,China;Department of Clinical Laboratory,Affiliated Hospital of Shaoxing University,Shaoxing 312000,Zhejiang,China)
出处
《中国现代医生》
2024年第23期30-34,共5页
China Modern Doctor
基金
浙江省教育厅一般科研项目(202145945)。
关键词
肝癌
尼克酰胺转甲基酶
细胞内活性氧
铁死亡
Liver cancer
Nicotinamide methyltransferase
Intracellular reactive oxygen species
Ferroptosis
