Cdt1和Mcm6作用片段在Y79细胞中的功能研究  

Analysis of Cdt1 and Mcm6 interaction domains in human retinoblastoma Y79 cells

作　　者：吴星[1] 王凤翔[1] 李朝辉[1] 

机构地区：[1]中国人民解放军总医院眼科,北京100853

出　　处：《中国斜视与小儿眼科杂志》2017年第1期1-4,共4页Chinese Journal of Strabismus & Pediatric Ophthalmology

摘　　要：目的探讨Cdc10依赖性转录因子1(Cdt1)和微小染色体维持蛋白6(Mcm6)对视网膜母细胞瘤Y79细胞增殖和凋亡的影响及作用机制。方法将人视网膜母细胞瘤细胞株Y79细胞分为五组,空质粒载体组、Cdt1(260-391)组,Cdt1(392-471)组,Mcm6(708-821)组,Mcm6(全长)组。脂质体转染法将各组质粒转染入Y79细胞;BrdU方法检测转染后的细胞增殖情况;流式细胞术检测细胞凋亡比例;染色质结合实验检测与染色质结合的内源性Mcm4、Mcm6和Cdc6。结果 Cdt1(392-471)组和Mcm6(708-821)组中Y79细胞的增殖能力低于其他三组,细胞凋亡率要高于其他三组,与染色质结合的内源性Mcm4和Mcm6减少。结论过度表达Cdt1和Mcm6的作用片段可以抑制Y79细胞的增殖,诱发细胞凋亡,其作用可能是抑制了内源性MCM蛋白复合体和DNA的结合。Objective To study the effect of Cdcl0 dependent transcriptl （Cdtl） and minichromosome mainte- nance6 （Mem6） on cell proliferation and apoptosis in a retinoblastoma cell line （Y79）, and explore the mechanism. Meth- ods The Y79 cells were divided into five groups which group cells were transfeeted with different plasmids： vector plas- mid,Cdtl（260-391） constructed plasmid, Cdt1（392-471） constructed plasmid,Mcm6（708-821） constructed plasmid and Mcm6 （Full length） constructed plasmid. BrdU incorporation assay and FACS analysis were performed to study the cell proliferation and apoptosis. Chromatin binding assay was used to detect the endogenous Mem4,Mcm6 and Cdc6. Results DNA replication was inhibited and apoptosis was induced in the cells transfected with Cdt1（392-471） constructed plasmid and the cell transfected with Mcm6（708-821） constructed plasmid. The endogenous Mcm4 and Mere6 were reduced in the two group cells. Conclusions Overexpression of Cdtl and Mere6 interaction domains in Y79 cells inhibit cell proliferation and induce cell apoptosis. The mechanism is probably to inhibit the DNA binding ability of MCM complex.

关 键 词：视网膜母细胞瘤 CDT1 Mcm6 

分 类 号：R739.7[医药卫生—肿瘤]