岩藻糖基转移酶8抑制剂2FF在多柔比星治疗肿瘤中的协同增效作用与机制  

Synergistic Effect and Mechanism of FUT8 Inhibitor 2FF With DOX for Cancer Treatment

作　　者：谢志东 章晓联[1,2] XIE Zhi-Dong;ZHANG Xiao-Lian(Hubei Key Laboratory of Allergy and Immune-related Diseases,Department of Immunology,Wuhan University Taikang Medical School(School of Basic Medical Sciences),Wuhan 430071,China;Department of Allergy,Zhongnan Hospital Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学泰康医学院(基础医学院)免疫学系,过敏及免疫相关疾病湖北省重点实验室,武汉430071 [2]武汉大学中南医院过敏科,武汉430071

出　　处：《生物化学与生物物理进展》2025年第2期478-486,共9页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(82230078,22077097);国家重点研发计划(2022YFA1303500)资助项目。

摘　　要：目的化疗是当前癌症治疗的重要手段之一,但化疗过程中多药耐药性的产生及副反应的发生严重限制了其应用,因此化疗常与其他药物或疗法联合应用。在已经鉴定的13个人类岩藻糖基转移酶(focosyltransferases,FUTs)中,FUT8(alpha-(1,6)-fucosyltransferase)是唯一负责核心岩藻糖基化的酶。核心岩藻糖基化在癌症的发生、转移及化疗耐药中有着重要作用,抑制FUT8是一种潜在逆转多药耐药的方法。本研究旨在探究FUT8的小分子抑制剂2FF(2-deoxy-2-fluoro-L-fucose)联合化疗药物多柔比星(doxorubicin,DOX)治疗恶性肿瘤的可行性。方法凝集素印迹实验检测2FF、DOX或二者联合对肿瘤细胞核心岩藻糖基化水平的影响。通过CCK-8法、Transwell侵袭实验探究2FF与DOX联合作用对人肝癌细胞HepG2及小鼠结肠癌细胞CT26细胞活性或侵袭的影响;通过流式细胞术检测2FF、DOX或二者联合对HepG2细胞表面程序性死亡受体配体1(programmed death-ligand 1,PD-L1)表达的影响;构建BABL/c小鼠结肠癌细胞皮下荷瘤模型以探究2FF与DOX联合作用对肿瘤生长的抑制作用。结果在本研究中,证实FUT8的抑制剂2FF能有效抑制HepG2及CT26细胞核心岩藻糖基化水平,而DOX可增强HepG2细胞核心岩藻糖基化水平,并且2FF可抑制这一作用。2FF能够有效增强HepG2及CT26细胞对DOX的敏感性。此外,2FF联合DOX可协同抑制HepG2细胞的侵袭能力。在体内水平,2FF联合DOX可增强对BALB/c小鼠CT26皮下荷瘤模型的疗效,但二者联合造成小鼠体重的下降。此外,DOX可引起HepG2细胞PD-L1表达上调,而2FF可削弱这一作用。结论FUT8抑制剂2FF可抑制DOX对肿瘤细胞的核心岩藻糖修饰和PD-L1表达的促进作用,并增强DOX的抗癌作用。Objective Chemotherapy is one of the important therapeutic approaches for cancer treatment.However,the emergence of multidrug resistance and side effects significantly limit its application.To address these challenges,chemotherapy is often combined with other drugs or therapies.Among the 13 human fucosyltransferases(FUTs)identified,FUT8(alpha-(1,6)-fucosyltransferase)is the only enzyme responsible for core fucosylation.Core fucosylation plays an important role in cancer occurrence,metastasis and chemotherapy resistance,making the suppression of FUT8 a potential strategy for reversing multidrug resistance.This study aims to evaluate the feasibility of combining the small molecule FUT8 inhibitor 2FF(2-deoxy-2-fluoro-L-fucose)with the clinical chemotherapeutic drug doxorubicin(DOX)for treating malignant tumors.Methods The human hepatocellular carcinoma cell line HepG2 and mouse colon cancer cell line CT26 cells were treated with 2FF,DOX or their combination and core fucosylation levels were assessed using Lectin Blot.HepG2 and CT26 cells were exposed to 50μmol/L 2FF for 72 h,followed by treatment with a gradient concentration of DOX for 24 h.Cell viability and IC50 values were determined via the CCK-8 assay.Transwell invasion assays were conducted to evaluate the combited effect of 2FF and DOX on the invasion ability of HepG2 cells.Flow cytometry was performed to analyze the impact of 2FF,DOX and their combination on membrane PD-L1 expression of HepG2 cells.To assess the in vivo effect,6 to 8 week old female BALB/c mice(20-25 g),were subcutaneously injected with 1×106 CT26 cells into the right axilla(four groups,six mice in each group).After the average tumor volume reached 100 mm3,mice were treated with DOX,2FF,their combination,or saline(mock group)every other day.DOX was administanted intraperitoneally(2 mg/kg),2FF intravenously(5 mg/kg),and the combination group,received the both treatment.Tumor size was measured every other day using a vernier caliper.Results This study demonstrated that DOX upregulates the c

关 键 词：核心岩藻糖基化 FUT8抑制剂 化疗 多柔比星 多药耐药 

分 类 号：R730.53[医药卫生—肿瘤]