家蚕细胞cyclinA基因的干涉对家蚕细胞增殖的影响  

RNA interference impact on silkworm cyclinA gene and cell proliferation

作　　者：刘丽华[1] 沈卫德[2] 李兵[2] 秦佳梅[1] 郁有兰 马鹏宇[1] 

机构地区：[1]通化师范学院生物系,吉林通化134000 [2]苏州大学基础医学与生物科学学院/现代丝绸国家工程实验室,江苏苏州215123 [3]吉林省抚松县第一中学,吉林白山134500

出　　处：《南京农业大学学报》2015年第1期168-171,共4页Journal of Nanjing Agricultural University

基　　金：吉林省科技厅项目(20130101100JC);吉林省教育厅项目(2013493)

摘　　要：[目的]研究小干扰RNA(small interfering RNA,siRNA)对家蚕卵巢细胞系(Bm N)细胞cyclin A基因表达及细胞周期的影响。[方法]通过脂质体转染试剂将针对cyclin A基因的特异性siRNA片段转入家蚕Bm N细胞,采用SYBR荧光定量PCR法检测cyclin A mRNA的表达,流式细胞技术检测细胞周期变化。[结果]转染siRNA-cyclin A可有效下调cyclin A基因的表达,转染48 h后,cyclin A mRNA表达量降低到对照的37.4%;流式细胞仪分析表明:与对照组相比,转染组G0/G1期细胞比例显著增加,细胞周期阻滞于G1期。[结论]靶向cyclin A基因的siRNA片段,通过下调cyclin A基因表达能有效抑制细胞增殖。[Objectives]The research was conducted to study small interfering RNA（ siRNA） the effects on silkworm ovary cell line（ Bm N） cyclin A gene expression and cell cycle. [Methods]Chemically synthesized siRNA fragments specific for cyclin A gene with liposomes was used to transfect Bm N cells,which were observed under confocal fluorescence microscopy,and mRNA expression was detected by real-time PCR,and cell cycle,by flow cytometry. [Results]With siRNA transfection efficiency into Bm N cells,and transfection siRNA-cyclin A after 48 h,silkworm cyclin A mRNA expression levels decreased 37.4%; flow cytometry showed that transfected group＇s transfer G0/ G1 phase cells rate increased,S phase and G2/ M phase of the cell ratio decreases. [Conclusions]The test show that imported siRNA fragments,cyclin A can be effectively inhibit,the cell cycle arrested in G1 phase,and inhibitted cell proliferation.

关 键 词：小干扰RNA(siRNA) 家蚕Bm N细胞 CYCLIN A基因 基因表达 细胞周期 

分 类 号：Q962[生物学—昆虫学]