TXNRD1在肝细胞癌中活性氧相关细胞凋亡导致多药耐药性中的作用  

Role of thioredoxin reductase 1 in multidrug resistance caused by reactive oxygen species-related cell apoptosis in hepatocellular carcinoma

作　　者：李大志 黄俊杰 郑树森[1] 章爱斌[2] LI Dazhi;HUANG Junjie;ZHENG Shusen;ZHANG Aibin(Key Laboratory of Multiple Organ Transplantation,The First Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 310000,China;Department of Hepatobiliary Surgery,The First Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 310000,China)

机构地区：[1]浙江大学医学院第一附属医院多器官联合移植重点实验室,杭州310000 [2]浙江大学医学院第一附属医院肝胆外科,杭州310000

出　　处：《临床肝胆病杂志》2022年第2期372-380,共9页Journal of Clinical Hepatology

摘　　要：目的耐药性是肝细胞癌(HCC)化疗失败的主要原因,硫氧还蛋白还原酶1(TXNRD1)作为活性氧(ROS)代谢的主要影响因素,已被证明与HCC患者的不良预后有关。本研究探究TXNRD1在HCC多药耐药机制中的作用。方法选取BEL-7402细胞系的BEL/FU细胞作为多重耐药细胞株。siRNA用来干预细胞TXNRD1的表达;实时定量PCR和蛋白质印迹检测TXNRD1的表达;CCK-8法检测和流式细胞术检测用来评估TXNRD1对肝癌细胞ROS蓄积、5-氟尿嘧啶(5-Fu)和多柔比星(DOX)耐药性以及体外凋亡的影响;异种肿瘤移植模型的构建用来研究金诺芬(AUR)在体内对细胞耐药性的影响。计量资料两组间比较采用独立样本t检验。结果BEL/FU作为多重耐药HCC细胞系表现出TXNRD1 mRNA与蛋白水平的高表达(P值均<0.05)。AUR作为TXNRD1的抑制剂,联合化疗药物5-FU及DOX处理与单一化疗药物处理相比,细胞克隆集落形成数目明显减少(P值均<0.01),凋亡比例明显增加(P值均<0.001)。N-乙酰半胱氨酸(NAC)作为ROS的清除剂,可以明显削弱siRNA敲低TXNRD1表达对BEL/FU细胞耐药性的影响,应用NAC可以有效降低干扰后细胞的凋亡比例(P值均<0.001)。动物实验也证实,5-Fu与AUR联合治疗相对于单一5-Fu治疗,裸鼠的肿瘤质量更轻(P<0.001),体积更小(P<0.001)。结论TXNRD1在HCC耐药中扮演重要角色,抑制其在细胞水平作用可有效改善耐药性,AUR作为TXNRD1抑制剂在HCC综合治疗中也具备极大的应用前景。Objective Drug resistance is the main cause of chemotherapy failure in hepatocellular carcinoma(HCC),and thioredoxin reductase 1(TXNRD1),as a major influencing factor for reactive oxygen species(ROS)metabolism,has been proven to be associated with the poor prognosis of patients with HCC.This study aims to explore the role of TXNRD1 in the mechanism of multidrug resistance in HCC.Methods BEL/FU cells in BEL-7402 cell line were selected as the multidrug-resistant cell line.The siRNA was used for the intervention of TXNRD1 expression;quantitative real-time PCR and Western blotting were used to measure the expression of TXNRD1;CCK-8 assay and flow cytometry were used to evaluate the effect of TXNRD1 on hepatocyte ROS accumulation,resistance to 5-fluorouracil(5-Fu)and doxorubicin(DOX),and apoptosis in vitro;a xenograft tumor model was established to investigate the effect of auranofin(AUR)on drug resistance in vivo.The two-independent-samples t test was used for comparison of continuous data between two groups.Results As a multidrug-resistant HCC cell line,BEL/Fu showed high mRNA and protein expression levels of TXNRD1(both P<0.05).Compared with 5-Fu or DOX treatment alone,the TXNRD1 inhibitor AUR combined with 5-Fu or DOX had had a significant reduction in the number of colony formation(P<0.01)and a significant increase in apoptosis ratio(P<0.001).The ROS scavenger N-acetylcysteine(NAC)significantly weakened the effect of TXNRD1 knockdown by siRNA on the drug resistance of BEL/Fu cells,and the application of NAC effectively reduced the apoptosis ratio of cells after siRNA interference(P<0.001).Animal experiments also confirmed that compared with the nude mice treated with 5-Fu alone,the nude mice treated with 5-Fu and AUR had a significantly lower tumor mass(P<0.001)and a significantly smaller tumor volume(P<0.001).Conclusion TXNRD1 plays an important role in the drug resistance of HCC,and inhibition of its level in cells can effectively improve drug resistance.As a TXNRD1 inhibitor,AUR has great application prospect

关 键 词：癌 肝细胞 活性氧 抗药性 肿瘤 硫氧还蛋白还原酶1 

分 类 号：R735.7[医药卫生—肿瘤]