异硫氰酸苄酯治疗未分化甲状腺癌的机制研究  

Mechanism of benzyl isothiocyanate in the treatment of undifferentiated thyroid cancer

作　　者：马春梅 韩朵 张惠英 杨磊[4] 李棣华[4] 张其成 王艳[6] 徐克[5] 贾强[1] 郑薇[1] 谭建[1] 孟召伟[1] Ma Chunmei;Han Duo;Zhang Huiying;Yang Lei;Li Dihua;Zhang Qicheng;Wang Yan;Xu Ke;Jia Qiang;Zheng Wei;Tan Jian;Meng Zhaowei(Department of Nuclear Medicine,Tianjin Key Lab of Functional Imaging and Tianjin Institute of Radiology,Tianjin Medical University General Hospital,Tianjin 300052;Fourth Department of Radiotherapy and Chemotherapy,NorthChina University of Science and Technology Affiliated Hospital,Tangshan 063000;Department of Imaging,NorthChina University of Science and Technology Affiliated Hospital,Tangshan 063000;Tianjin Key Laboratory of AcuteAbdomen Disease Associated Organ Injury and ITCWM Repair,Institute of Acute Abdominal Diseases,Tianjin Nankai Hospital,Tianjin 300100;Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment,TianjinLung Cancer Institute,Tianjin Medical University General Hospital,Tianjin 300052;State Key Laboratory ofComponent-Based Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617;Department of Imaging,Affiliated Hospital of North China University of Science andTechnology,Tangshan 063000,China)

机构地区：[1]天津医科大学总医院核医学科,天津市功能影像重点实验室和天津市影像医学研究所,天津300052 [2]华北理工大学附属医院肿瘤放化疗四科,唐山063000 [3]华北理工大学附属医院影像科,唐山063000 [4]天津市急腹症相关器官损伤及ITCWM修复重点实验室,急性腹部疾病研究所,天津市南开医院,天津律300102 [5]天津市肺癌转移与肿瘤微环境重点实验室,天津市肺癌研究所,天津医科大学总医院,天津300052 [6]中药成分国家重点实验室,天津中医药大学,天津301617 [7]华北理工大学附属医院影像科工作,唐山063000

出　　处：《中华内分泌代谢杂志》2024年第11期966-977,共12页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金(81971650);天津市科委基金项目(21JCYBJC01820);天津市医学重点学科(专科)建设项目(TJYXZDXK-001A)。

摘　　要：目的研究异硫氰酸苄酯(benzyl isothiocyanate, BITC)治疗未分化甲状腺癌(anaplastic thyroid cancer, ATC)的机制。方法网络药理学分析:BITC和ATC关键靶点筛选, 并进行GO和KEGG富集分析。验证:运用AutoDock软件对BITC及ATC关键靶点进行对接;细胞实验:首先, BITC作用于2种ATC细胞株8505C和CAL-62, 流式细胞术分析细胞凋亡情况, 然后自噬抑制剂硫酸羟基羟氯喹(HCQ)和3-甲基腺嘌呤(3MA)分别与BITC联合应用, 实时荧光定量PCR检测LC3B基因水平及Western印迹法检测NF-κB、LC3BⅡ、Beclin-1、Bcl-2表达量;动物实验:CAL-62构建小鼠肿瘤模型, 分别用腹腔注射BITC(100 mg/kg)和生理盐水治疗, 隔天给药, 共21 d, 肿瘤组织免疫印迹检测LC3B Ⅱ、Bcl-2、Beclin-1及NF-κB的表达。结果共筛选出10个关键靶点, 对接结合能均小于≤-4.0 kcal/mol;KEGG显示关键靶基因主要富集于NF-κB信号通路、凋亡等信号通路。BITC对2种ATC细胞株8505C和CAL-62有抑制作用, IC50值分别为27.56和28.30 μmol/L, NF-κB、Beclin-1及Bcl-2的表达降低, 而LC3B Ⅱ表达升高, LC3B基因表达水平明显升高。3MA与BITC联合应用, 抑制细胞作用增强, 同时增加LC3B Ⅱ的表达;HCQ增加LC3B Ⅱ表达, 但并未增强抑制细胞活力作用。小鼠肿瘤模型中, 与对照组相比治疗组LC3B Ⅱ表达增加, Bcl-2、Beclin-1及NF-κB表达降低。结论 BITC在体外和体内均抑制ATC细胞的生长, 破坏ATC细胞的自噬降解, 抑制NF-κB通路。Objective To investigate the mechanism of benzyl isothiocyanate(BITC)in the treatment of anaplastic thyroid cancer(ATC).Methods Using network pharmacological analysis,key targets of BITC and ATC were screened,followed by CO and KEGG enrichment analysis.In order to validate the findings,AutoDock software was used to dock BITC and ATC key targets.BITC was applied to two ATC cell lines(8505C and CAL-62).Flow cytometry was used to analyze cell apoptosis.Autophagy inhibitors hydroxychloroquine sulfate(HCQ)and 3-methyladenine(3MA)were used in combination with BITC.Real-time quantitative PCR was conducted to detect the gene level of LC3B,while Western blotting was utilized to examine the expression of NF-κB,LC3B II,Beclin-1,and Bcl-2.In animal experiments,a mouse tumor model was constructed using CAL-62 cells,treated with intraperitoneal injections of BITC(100 mg/kg)and normal saline respectively,administered every other day for a total of 21 days.Immunoblotting of tumor tissue was performed to detect the expression of LC3B II,Bcl-2,Beclin-1,and NF-κB.Results A total of 10 key targets with binding energies≤-4.0 kcal/mol were identified.KECG analysis showed that these genes are mainly involved in NF-κB signaling pathway and apoptosis.BITC inhibited ATC cells with IC50 values of 27.56μmol/L for 8505C and 28.30μmol/L for CAL-62.The expression levels of NF-κB,Beclin-1,and Bcl-2 decreased,while LC3B II and LC3B gene expression increased.Combining 3MA with BITC enhanced cell inhibition LC3B I expression.HCQ increased LC3B I expression without enhancing cell and viability inhibition.In the mouse tumor model,compared to the control group,the treatment group had higher LC3B II and lower Bcl-2,Beclin-1,and NF-κB levels.Conclusion BITC could inhibit the growth of ATC cells in vitro and in vivo,disrupt the autophagy degradation,and inhibit the NF-κB pathway.

关 键 词：异硫氰酸苄酯 未分化甲状腺癌 自噬 NF-ΚB信号通路 细胞凋亡 

分 类 号：R736.1[医药卫生—肿瘤]