论文部分内容阅读
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。
Using RNAdraw and Mfold bioinformatics software to predict the secondary structure of cyclin D1 mRNA, cyclin D1-DRz targeting cyclin D1 was designed and transfected into tumor Cells u251 and HeLa. The results of reverse transcription-polymerase chain reaction (RT-PCR) showed that cyclin Dl-DRz significantly inhibited the expression of cyclin D1 gene in u251 and HeLa cells and inhibited the expression of cyclin E1, cyclin A1 and cyclin B1 Such as decreased expression. Flow cytometry analysis of cell cycle changes before and after transfection showed tumor cell arrest in the G0 / G1 phase increased. This experiment established a screening platform for the target site of DNAzyme and confirmed that cyclin Dl-DRz can effectively interfere with the process of tumor cells in the cell cycle.