miR-449b和miR-34c对卵巢癌细胞SKOV3-ipl周期相关蛋白的下调及细胞周期阻滞作用  

miR-449b and miR-34c on inducing down-regulation of cell cycle-related proteins and cycle arrests in SKOV3-ipl cell,an ovarian cancer cell line

作　　者：马丽萍[1] 李娜[1] 何湘君[1] 张旗[1] 

机构地区：[1]北京大学人民医院临床分子生物学研究所,北京100044

出　　处：《北京大学学报（医学版）》2011年第1期129-133,共5页Journal of Peking University:Health Sciences

摘　　要：目的：基于miR-449和miR-34在p53突变的卵巢癌细胞系SKOV3和SKOV3-ipl的表达差异,研究探讨这些miRNA对肿瘤细胞生长、细胞周期的影响及靶基因的表达变化。方法：通过反转录实时定量PCR方法测定miR-449a/b和miR-34b,c在SKOV3和SKOV3-ipl的表达,通过转染使它们在极低表达的SKOV3-ipl中获得表达。用MTS方法测定细胞生长率变化、流式细胞术检测细胞周期改变、Western blot检测细胞周期相关蛋白表达。结果：miR-449b和miR-34c使SKOV3-ipl黏附性下降28%~34%,细胞周期阻滞：G1期细胞数量分别增加15.62%和15.71%;S期细胞数量分别减少15.96%和16.56%。细胞周期相关蛋白CDK6和CDC25A均表达下调,miR-449b使CDK6减少39%、CDC25A减少22%;miR-34c使CDK6减少49%、CDC25A减少32%;而miR-449b与miR-34c共同作用后,下调作用更明显,CDK6减少69%,CDC25A减少86%,CyclinA减少59%。结论：miR-449b和miR-34c使高恶性卵巢癌细胞系SKOV3-ipl发生细胞周期阻滞,CDK6、CDC25A和CyclinA表达均下调。Objective：To investigate the effects of miR-449 and miR-34 on cell growth,cell cycle and target gene expression based on these miRNA different expressions in ovarian cancer cell lines SKOV3 and SKOV3-ipl both with mutation of p53.Methods： The expressions of miR-449a/b and miR-34b,c in SKOV3 and SKOV3-ipl were detected by RT-PCR.miR-449a,b and miR-34b,c were ectopically expressed by transfection of SKOV3-ipl.The cell growth rate was assayed by MTS method.The changes of cell cycle were measured by FCM.The changes of expression of cell cycle related proteins were detected by Western blot.Results： Ectopic expression of miR-449b and miR-34c resulted in lowered adhesion activities by 28%-34%,and in cell cycle arrests with increased cell number of 15.62% and 15.71% in G1 and with decreased cell number of 15.96% and 16.56% in S.Cell cycle related proteins CDK6 and CDC25A were down-regulated.The decreases of CDK6 and CDC25A by miR-449b were 39% and 22% respecyively;49% and 32% by miR-34c respectively.The more decreases were seen in co-action by miR-449b and miR-34c with decreases of 69% in CDK6,86% in CDC25A,and 59% in CyclinA.Conclusion： miR-449b and miR-34c resulted in cell cycle arrests and down-regulation of CDK6,CDC25A and CyclinA in high malignant ovarian cancer cell line SKOV3-ipl.

关 键 词：卵巢肿瘤 细胞周期蛋白类 微RNAS 

分 类 号：R349.7[医药卫生—基础医学]