摘要
目的:探讨三结构域蛋白59(tripartite-motif protein 59,TRIM59)对于人肝细胞肝癌(hepatocellular carcinoma, HCC)细胞增殖与迁移的影响以及波形蛋白(vimentin, VIM)作为其中潜在调控靶点的相关性研究。方法:通过细胞培养和细胞转染得到所需要的人HCC细胞,采用定量实时聚合酶链反应(qRT-PCR)方法检测其中TRIM59 mRNA的表达水平并通过蛋白印迹法(Western blot)检测TRIM59蛋白质的表达水平;采用慢病毒介导的基因沉默效应与TRIM59过表达质粒进行转染介导的基因过表达效应调控TRIM59基因的表达,通过集落形成试验检测TRIM59处于不同表达水平时的肝癌细胞系的增殖能力差别;通过细胞划痕试验检测TRIM59基因处于不同表达水平时HCC细胞的迁移能力差异;并通过考马斯亮蓝法(Bradford法)检测TRIM59基因处于不同表达水平时,VIM的表达情况,通过统计学方法研究TRIM59及VIM表达的相关性。结果:通过定量qRT-PCR与Western blot检测,在所选取的HCC细胞系中,TRIM59 mRNA及TRIM59蛋白质的表达量均高于普通肝细胞(P均<0.01)。细胞集落形成实验显示,在所选的肝癌细胞中,TRIM59的表达与HCC的细胞集落形成数量呈正相关(P均<0.01)。细胞划痕试验显示,TRIM59的表达与HCC的细胞迁移能力呈正相关(P均<0.05)。Bradford法检测显示,在所选取的HCC细胞系中,TRIM59蛋白质的表达与VIM的表达呈正相关(P均<0.01)。结论:TRIM59促进人HCC细胞的增殖与迁移,可能是通过调控VIM表达而完成,可为HCC的靶向治疗提供新的靶点。
Objective:To investigate the influence of TRIM59 on proliferation and migration of human hepatocellular carcinoma(HCC)and the correlation of vimentin as a potential regulatory target.Methods:The required human hepatocellular carcinoma cells were obtained by cell culture and transfection.The expression level of TRIM59 mRNA was detected by qRT-PCR and the expression level of TRIM59 protein was detected by Western blot.Using slow virus mediated gene silencing effect with TRIM59 overexpression plasmid transfection mediated gene overexpression effect regulated TRIM59 expression.By colony formation test,cell proliferation ability difference of TRIM59 in different expression levels of liver cancer was detected.Cell scratch test was used to detect the difference of migration ability of HCC at different expression levels of TRIM59.Through the Bradford method,TRIM59 gene expression in different levels,the expression of vimentin was detected.Through statistical methods the correlation between TRIM59 and VIM protein in HCC proliferation was analysed.Results:By quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot analysis,the expression levels of TRIM59 mRNA and TRIM59 protein in the selected HCC were higher than that of ordinary liver cells(all P<0.01).The cell aggregation experiment showed that in selected HCC cells,the expression intensity of TRIM59 was positively correlated with the number of HCC cell colonies(all P<0.01).Cell scratch test showed that the expression intensity of TRIM59 was positively correlated with the migration ability of HCC cells(all P<0.05).Bradford analysis showed that the expression intensity of TRIM59 protein was positively correlated with the expression intensity of vimentin in selected HCC(all P<0.01).Conclusion:TRIM59 promotes the proliferation and migration of human HCC,which may be regulated by regulating VIM,providing a new target for targeted therapy of HCC.
作者
刘德权
孙纲
李校冲
刘晗
LIU Dequan;SUN Gang;LI Xiaochong;LIU Han(The No.967 Hospital of PLA Joint Logistics Support Force,Postgraduate Culture Base of Jinzhou Medical University,Liaoning Dalian 116021,China;The No.967 Hospital of PLA Joint Logistics Support Force,Liaoning Dalian 116021,China;The No.967 Hospital of PLA Joint Logistics Support Force,Postgraduate Culture Base of Dalian Medical University,Liaoning Dalian 116021,China)
出处
《现代肿瘤医学》
CAS
北大核心
2021年第19期3341-3346,共6页
Journal of Modern Oncology
基金
辽宁省自然科学基金(编号:20180550603)。
关键词
TRIM59
肝细胞肝癌
VIM
增殖
迁移
TRIM59
hepatocellular carcinoma
vimentin
proliferation
migration
