STARD7蛋白新型小分子抑制剂Lasiodin激活NRF2/EGR1通路诱导三阴乳腺癌铁死亡  

The novel inhibitor of STARD7 induces ferroptosis through the NRF2/EGR1 pathway in TNBC

作　　者：李雪[1] 苏小涵 曾姣 梁婷婷 屈鹏 刘俊 王雅丽[2] 侯令密[2] 郭晓兰[1] 梁骑[1] LI Xue;SU Xiaohan;ZENG Jiao;LIANG Tingting;QU Peng;LIU Jun;WANG Yali;HOU Lingmi;GUO Xiaolan;LIANG Qi(Department of Clinical Laboratory,Affiliated Hospital of North Sich-uan Medical College,Nanchong 637000,Sichuan,China;Department of Breast and Thyroid Surgery,Affiliated Hospital of North Sich-uan Medical College,Nanchong 637000,Sichuan,China;School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 404100,China)

机构地区：[1]川北医学院附属医院检验科,南充637000 [2]川北医学院附属医院甲状腺乳腺外科,南充637000 [3]重庆理工大学药学与生物工程学院,重庆404100

出　　处：《医学研究与战创伤救治》2024年第5期462-471,共10页Journal of Medical Research & Combat Trauma Care

基　　金：四川省自然科学基金(2022NSFSC0775);川北医学院博士科研启动基金(CBY21-QD07)。

摘　　要：目的探讨StAR相关脂质转移结构域包含7(STARD7)在三阴乳腺癌(TNBC)中的表达情况及其与预后的相关性。高通量筛选STARD7的小分子抑制剂,研究其小分子抑制剂对TNBC的影响及潜在分子机制。方法生物信息学分析STARD7在各亚型乳腺癌中的表达情况以及与预后的相关性。Westernblot和免疫组化检测细胞系以及临床TNBC样本中STARD7的表达情况。以STARD7为靶点,通过分子对接、体外蛋白纯化和体外分子互作实验,筛选并鉴定STARD7的小分子抑制剂。CCK8测定对照组(DMSO组)及1、2、5、10、20μmol/L浓度Lasiodin处理24 h后的TNBC细胞的存活率,确定最佳药物作用浓度。台盼蓝和YO-PRO-1染色测定各组细胞死亡率,Westernblot测定各组凋亡、铁死亡相关蛋白表达;RT-qPCR及Western blot分别测定NRF2、HO-1、KEAP1、EGR1的mRNA和蛋白表达。构建TNBC移植瘤模型并随机分为对照组、紫杉醇组(10 mg/kg)和Lasiodin组(10 mg/kg),分别处理后测定肿瘤体积、裸鼠体重,HE染色评价生物安全性。结果生物信息学分析及体外实验表明STARD7在TNBC组织及细胞中高表达,且与患者预后负相关(P<0.05);高通量筛选并鉴定出Lasiodin是STARD7的小分子抑制剂;Lasiodin对STARD7的抑制可能通过调控下游NRF2/EGR1通路相关蛋白的表达促使TNBC细胞发生铁死亡,同时伴有凋亡和坏死。体内实验证明Lasiodin能够抑制肿瘤生长(P<0.05),且未观察到明显的不良反应。结论STARD7可能在TNBC的进展中发挥重要的调控作用,Lasiodin为其小分子抑制剂,提示STARD7和Lasiodin分别可能成为治疗TNBC的潜在靶点和新型药物。Objective To investigate the expression of StAR-related lipid transfer domain7(STARD7)in triple-negative breast cancer(TNBC)and its correlation with prognosis.To perform highthroughput screening for small molecule inhibitors of STARD7,and to study their effects on TNBC and their potential molecular mechanisms.Methods Bioinformatics analysis of the expression of STARD7 in breast cancer and its correlation with prognosis.Western blot(WB)and immunohistochemistry experiments were used to detect the expression of STARD7 in cell lines and clinical TNBC samples.Targeting STARD7,molecular docking,in vitro protein purification,and in vitro molecular interaction experiments were conducted to screen and identify small molecule inhibitors of STARD7.The CCK8 assay was used to determine the survival rate of TNBC cells after 24 hours of treatment with control group(DMSO)and Lasiodin at concentrations of 1,2,5,10,and 20μmol/L to identify the optimal drug concentration.The cell death ratio of each group was determined by trypan blue and YO-PRO-1 staining,and the expression of apoptosis-and ferroptosis-related proteins was measured by Western blot.The expressions of mRNAs and proteins of NRF2,HO-1,KEAP1,and EGR1 were measured by RT-qPCR and Western blot,respectively.A TNBC xenograft model was established and randomly divided into control group,paclitaxel group(10 mg/kg),and Lasiodin group(10 mg/kg).After treatment,tumor volume and nude mouse body weight were measured,and biosafety was evalu⁃ated through HE staining.Results Bioinformatics analysis and in vitro experiments indicated that STARD7 was highly expressed in TNBC tissues and cells,and its high expression was negatively correlated with patient prognosis(P<0.05).High-throughput screening identified Lasiodin as a small molecule inhibitor of STARD7.The inhibition of STARD7 by Lasiodin may induce ferroptosis in TN⁃BC cells by regulating the expression of proteins related to the downstream NRF2/EGR1 pathway,along with apoptosis and necrosis.In vivo experiments demonstrated

关 键 词：STARD7 毛栲利素 三阴乳腺癌 分子对接 小分子抑制剂 铁死亡 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]