基于“睡美人”转座系统观察OGT抑制剂HLY838对肝癌的治疗作用  

Therapeutic effect of the O-linked N-acetylglucosamine transferase inhibitor HLY838 on liver cancer:a study based on the Sleeping Beauty transposition system

作　　者：梁惠钧 黄露义 汪凯[1] 唐霓[1] Liang Huijun;Huang Luyi;Wang Kai;Tang Ni(Key Laboratory of Molecular Biology of Infectious Diseases Designated by the Chinese Ministry of Education,Chongqing Medical University)

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《重庆医科大学学报》2025年第1期30-36,共7页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:82272975)。

摘　　要：目的:利用“睡美人”转座系统和快速大容量尾静脉注射法,建立肝癌动物模型,并观察O-连接N-乙酰氨基葡萄糖转移酶(O-linked N-acetylglucosamine transferase,OGT)抑制剂(R)-3-(2-甲氧基苯基)-1-(噻吩-2-基甲基)哌嗪-2,5-二酮[(R)-3-(2methoxyphenyl)-1-(thiophen-2-ylmethyl)piperazine-2,5dione,HLY838]对肝癌的治疗效果。方法:将6~8周龄雄性C57BL/6J小鼠随机分为3组,分别将插入神经母细胞瘤ras病毒同源物(neuroblastoma RAS viraloncogene homolog,NRAS)的NRASV12转座子质粒、表达β-连环蛋白(β-catenin)基因转座子质粒、表达Yes相关蛋白(Yes-associated protein,YAP)基因的转座子质粒pT3-EF1aH Yap S127A与转座酶SB100质粒的混合溶液经尾静脉快速注射至小鼠体内,饲养8~16周后处死小鼠,观察肝脏成瘤情况以及肝脏病理学变化。肝癌模型建成后,腹腔注射HLY838,观察对肝癌的治疗效果。结果:高压注射质粒8~16周后,小鼠肝脏体积增大,表面出现颗粒状、结节样改变;苏木精-伊红染色(hematoxylin-eosin staining,HE)染色显示肝组织结构紊乱,出现多个肿瘤结节。pT3-EF1aH YAP S127A组成瘤率高、时间短,但肿瘤较小;pT/CAGGS-NrasV组肿瘤结节大小适中,但成瘤时间较长;pT3-N90-β-catenin组肿瘤主要为肝细胞癌。HLY838治疗组小鼠肝肿瘤结节数变少,小鼠肝肿瘤负担减轻。结论:应用水动力大容量尾静脉注射质粒的方法可快速建立小鼠肝癌模型,成功率较高。OGT抑制剂HLY838对小鼠肝癌有较好的抗肿瘤效果。Objective:To establish an animal model of liver cancer by rapid high-volume tail vein injection,and to investigate the therapeutic effect of O-linked N-acetylglucosamine transferase inhibitor(R)-3-(2methoxyphenyl)-1-(thiophen-2-ylmethyl)piperazine-2,5dione(HLY838)on liver cancer based on the Sleeping Beauty(SB)transposition system.Methods:Male C57BL/6J mice,aged 6-8 weeks,were randomly divided into three groups and were given the NRASV12 transposon plasmid,theβ-catenin geneexpressing transposon plasmid,or the YAP gene-expressing transposon plasmid pT3-EF1aH Yap S127A,mixed with the transposase SB100 plasmid,through rapid high-volume tail vein injection.The mice were sacrificed after 8-16 weeks of feeding to observe tumor formation and liver pathological changes.After the establishment of the liver cancer model,HLY838 was injected intraperitoneally to investigate its therapeutic effect on liver cancer.Results:After high-pressure injection of the plasmids for 8-16 weeks,there was an increase in liver volume,with granular and nodular changes on the surface of the liver,and HE staining showed disordered structure of the liver and the presence of multiple tumor nodules.The pT3-EF1aH YAP S127A group had a higher tumor formation rate and a shorter duration,but with a relatively small tumor size;the pT/CAGGS-NrasV group had moderately sized tumor nodules,but with a relative long time for tumor formation;the pT3-N90-β-catenin group mainly had hepatocellular carcinoma.The HLY838 treatment group had reductions in the number of liver tumor nodules and the burden of liver tumor.Conclusion:The method of hydrodynamic high-volume tail vein injection can be used to establish a mouse model of liver cancer rapidly,with a relatively high success rate.The OGT inhibitor HLY838 has a good antitumor effect on liver cancer in mice.

关 键 词：肝肿瘤 动物模型 “睡美人”转座系统 水动力尾静脉注射 (R)-3-(2-甲氧基苯基)-1-(噻吩-2-基甲基)哌嗪-2 5-二酮 

分 类 号：R735.7[医药卫生—肿瘤]