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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 investigate the effect and mechanism of Cdc10-dependent transcription factor 1 (Cdt1) and Mcm6 on proliferation and apoptosis of retinoblastoma Y79 cells. Methods Human retinoblastoma cell line Y79 cells were divided into five groups: empty plasmid vector group, Cdt1 (260-391) group, Cdt1 (392-471) group, Mcm6 (708-821) group. The plasmids were transfected into Y79 cells by Lipofectamine 2000 transfection method; the proliferation of transfected cells was detected by BrdU method; the apoptosis ratio was detected by flow cytometry; the chromatin binding assay was used to detect the endogenous Mcm4, Mcm6 and Cdc6. Results The proliferation of Y79 cells in Cdt1 (392-471) and Mcm6 (708-821) groups was lower than that in the other three groups. The apoptotic rate of Y79 cells was higher than that of the other three groups. The endogenous Mcm4 and Mcm6 cut back. Conclusion The overexpression of Cdt1 and Mcm6 can inhibit the proliferation and induce the apoptosis of Y79 cells, which may be due to the inhibition of the binding of endogenous MCM protein complexes to DNA.