药物重定位技术筛选双硫仑抑制骨肉瘤生长的靶点  

Screening the target of disulfiram for inhibiting the growth of osteosarcoma by drug repositioning technique

作　　者：付东 徐平 宁波 王达辉 Fu Dong;Xu Ping;Ning Bo;Wang Dahui(Department of Orthopedics,Children′s Hospital of Fudan University,National Children′s Medical Centre,Shanghai 201102,China)

机构地区：[1]国家儿童医学中心/复旦大学附属儿科医院骨科,上海201102

出　　处：《中华转移性肿瘤杂志》2023年第4期403-409,共7页Chinese Journal of Metastatic Cancer

基　　金：上海市扬帆计划项目(21YF1403500)。

摘　　要：目的用药物重定位技术筛选双硫仑对骨肉瘤裸鼠移植瘤生长的靶点,为骨肉瘤患者治疗提供依据。方法在最新的小分子抗癌化合物库中筛选出显著抑制骨肉瘤生长的小分子化合物——双硫仑。先用143B骨肉瘤进行细胞实验,然后构建裸鼠骨肉瘤模型并验证双硫仑对骨肉瘤裸鼠移植瘤生长曲线和生存率的影响。结果双硫仑能显著抑制143B骨肉瘤细胞活力和增殖。骨肉瘤裸鼠移植瘤实验结果证实双硫仑降低了骨肉瘤体积,提高了生存率。进一步研究发现143B细胞中组蛋白去甲基转移酶5的mRNA以及蛋白质水平在给予双硫仑刺激后均显著降低。免疫荧光结果显示双硫仑处理143B细胞后,KDM5A荧光强度显著降低。药物亲和力响应目标稳定性和微量热泳动结果显示双硫仑和KDM5A有着直接相互作用关系。结论双硫仑通过下调去甲基转移酶5的表达抑制了骨肉瘤裸鼠移植瘤生长,可能成为未来临床骨肉瘤治疗的有效药物。Objective To screen the target of disulfiram for inhibiting the growth of transplantable osteosarcoma in nude mice with drug repositioning technology,and to provide a basis for the treatment of osteosarcoma patients.Methods The small molecule compound,disulfiram,which significantly inhibited the growth of osteosarcoma was selected from the latest library of small molecule anticancer compounds.Firstly,cell experiments were performed with 143B osteosarcoma.Then,the nude model of osteosarcoma was constructed to verify the effects of disulfiram on the growth curve and survival rate of transplantable osteosarcoma.Results Disulfiram could significantly inhibited cell viability and proliferation of 143B osteosarcoma cells.The results of in vivo experiment confirmed that disulfiram reduced the volume of osteosarcoma in mice and increased the survival rate.The mRNA and protein levels of histone demethyltransferase 5 in 143B cells were significantly reduced after being stimulated with disulfiram.The KDM5A fluorescence intensity was decreased in 143B cells by Immunofluorescence following treatment with disulfiram.Drug affinity response to target stability and microscale thermophoresis results indicated a direct interaction between disulfiram and KDM5A.Conclusions Disulfiram inhibits the growth and proliferation of osteosarcoma by down-regulating the expression of desmethyltransferase 5,and may be used as an effective drug for the clinical treatment of osteosarcoma.

关 键 词：药物重定位 双硫仑 去甲基转移酶5 骨肉瘤 

分 类 号：R738[医药卫生—肿瘤]