靶向cyclin D1的脱氧核酶对肿瘤细胞周期的影响  

Influence of DNAzymes against Cyclin D1 in Tumor Cell Cycle

作　　者：李丹[1] 朱俊袆[1] 周雅彦[1] 刘选明[1] 

机构地区：[1]湖南大学生命科学与技术研究院,长沙410082

出　　处：《生物医学工程学杂志》2009年第2期374-378,384,共6页Journal of Biomedical Engineering

基　　金：国家高校“985工程”创新平台项目资助;湖南大学自然科学基金资助项目(Y00003)

摘　　要：应用RNAdraw和Mfold生物信息学软件进行细胞周期蛋白D1(cyclin D1)mRNA的二级结构预测,设计靶向cyclin D1的10-23型脱氧核酶(cyclin Dl-DRz),将其转染至肿瘤细胞u251和HeLa中。逆转录-聚合酶链反应技术(RT-PCR)检测发现,cyclin Dl-DRz能明显抑制u251、HeLa细胞中cyclin D1基因的表达,并使细胞周期蛋白E1、细胞周期蛋白A1和细胞周期蛋白B1等表达下降。流式细胞仪技术分析转染前后细胞周期的变化,显示肿瘤细胞阻滞在G0/Gl期的比例增加。本实验建立了脱氧核酶靶位点的筛选平台,并证实cyclin Dl-DRz能有效干扰肿瘤细胞在细胞周期中的进程。In this study, DNAzymes against cyclin D1 （cyclin D1-DRz） were designed according to the secondary structure of cyclin D1 mRNA which was computed with RNAdraw and Mfold. Cyclin D1-DRz were transfected into tumor cell line u251 and HeLa by oligofectamine. The expression of cyclin D1 was detected by RT-PCR. It was shown that the expression of cyclin D1 gene was suppressed obviously, and the expressions of other cell-cycle related genes such as cyclin El, cyclin A1 and cyclin B1 were also declined. The cell cycle analysis of tumor cells tansfected with cyclin D1-DRz revealed an arrestment in the G0/G1 phase. In conclusion,the approach is effective and feasible for designing DNAzyme. Cyclin D1-DRz is useful for interfering with the cell cycle procession of tumor cells.

关 键 词：RNAdraw Mfold 细胞周期蛋白 D1 脱氧核酶 细胞周期 

分 类 号：R73-3[医药卫生—肿瘤]