siRNA-CyclinE对家蚕细胞增殖的影响  

Influence of siRNA-CyclinE on proliferation of BmN cells

作　　者：刘丽华[1] 沈卫德[2,3] 李兵[2,3] 王文兵[4] 

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,江苏苏州215123 [3]现代丝绸国家工程实验室,江苏苏州215123 [4]江苏大学生命科学研究院,江苏镇江212001

出　　处：《西北农林科技大学学报（自然科学版）》2013年第4期21-25,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(31072086)

摘　　要：【目的】研究小干扰RNA(Small interfering RNA,siRNA)对家蚕卵巢细胞系(BmN)细胞CyclinE基因表达及细胞周期的影响。【方法】合成针对CyclinE基因1 252和568位点的siRNA片段,并利用脂质体法转染家蚕BmN细胞,于共聚焦荧光显微镜下观察转染情况;应用实时荧光定量PCR法检测CyclinE mRNA的表达情况,流式细胞仪检测细胞周期。【结果】siRNA对家蚕BmN细胞的转染效率达80%左右。转染1252-siRNA-CyclinE和568-siRNA-CyclinE 48h后,分别使家蚕BmN细胞CyclinE mRNA表达量降低了68%和90%;流式细胞仪检测结果显示,2个转染组G0/G1期细胞比例显著增大,S期和G2/M期细胞比例显著减小。【结论】在家蚕BmN细胞中导入针对CyclinE的siRNA,可有效抑制CyclinE基因的表达,使细胞周期阻滞于G1期,进而抑制细胞增殖。[Objective] This study aimed to study the influence of small interfering RNA on CyclinE gene expression of cell and cell cycle in Bombyx rnori ovarian cell lines （BmN）. [Method] siRNA for CyclinE gene fragments at the site of 1 252 and 568 was chemically synthesized before being transfected into BmN cells by lipofectin. Transfection efficiency was observed using confocal fluorescence microscope and then the CyclinE mRNA expression was determined by real-time fluorescent quantitative PCR method. At last,the cell cycle was detected by flow cytometry. [Result] BraN cells transfection efficiency of siRNA could reach to 80%. 48 h after the transfection of 1252-siRNA-CyclinE and 568-siRNA-CyclinE,BmN cells CyclinE mRNA expression were reduced by 68% and 90% respectively. Flow cytometry results in the 2 transfection groups showed that the G0/G1 phase cells ratio increased,while the S and G2/M phase cell ratios decreased. [Conclusion] The siRNA against CyclinE was imported into BraN cells,and showed effectively inhibition to CyclinE gene expression, and bogged down cell cycle in G1 phase, so as to inhibit cell proliferation.

关 键 词：家蚕BmN细胞 CyclinE基因 RNA干扰 细胞增殖 细胞周期 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]