siRNA沉默DNA甲基化转移酶1基因干扰胎牛成纤维细胞增殖和凋亡(英文)  

Interference of DNA Methyltransferase 1 Gene Silenced by siRNA on Fetal Bovine Fibroblast Cell Proliferation and Apoptosis

作　　者：于孟飞 王文璐 易建明 Yu Mengfei;Wang Wennu;Yi Jianming(Institute tor Medical Biology and Hubei Provincial Key Laboratory tor Protection and Application of Special Plant Gennplasm in the Wuling Area of China,College of Lite Sciences,South-Central University tor Nationalities,Wuhan 430074,China;Key Laboratory of Agricultural Animal Genetic,Breeding and Reproduction tor Minisntry of Education,College of Animal Science,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]中南民族大学生命科学学院,医学生物研究所&武陵山区特色资源植物种质保护与利用湖北省重点实验室,武汉430074 [2]华中农业大学动物科技学院,农业动物遗传育种与繁殖教育部重点实验室,武汉430074

出　　处：《中南民族大学学报（自然科学版）》2018年第2期24-30,共7页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：国家自然科学基金青年资助项目(81400252);中央高校基本科研业务费专项资金资助项目(2013PY130)

摘　　要：为阐明Dnmt1-siRNA对胎牛成纤维细胞(FBFCs)的影响,设计了3种针对Dnmt1的siRNA来转染FBFCs.结果显示:Dnmt1-siRNA3转染FBFCs 24,48,72 h后,Dnmt1 mRNA表达水平显著降低(P<0.01).在转染后48 h,siRNA3对Dnmt1抑制效率达到近80%,Dnmt1-siRNA3处理的FBFCs增殖也受到显著影响,FBFC细胞活力下降、处于G0/G1过渡期细胞数量增多(P<0.05).但Dnmt1-siRNA3组FBFC细胞的凋亡率也显著增加(P<0.05).说明通过Dnmt1特异性siRNA能够有效地敲低FBFC中的Dnmt1 mRNA表达量,由于细胞周期分布变化,siRNA处理后FBFC作为核供体能提高SCNT的效率.To determine the effects of siRNAs targeting Dnmt1 on fetal bovine fibroblast cells（ FBFCs）,three siRNAs against Dnmt1 were designed for transfection into FBFCs. The results showed that Dnmt1-siRNA3 significantly reduced the mRNA expression level at 24,48 and 72 h（ P〈0.01）. At 48 h post-transfection,the inhibitory efficacy of siRNA3 against Dnmt1 reached nearly 80%.Dnmt1-siRNA3 treatment of FBFCs also significantly inhibited their proliferation,reduced cell viability and arrested more cells in the G0/G1 transition（ P〈0. 05）. However,Dnmt1-siRNA3 increased the number of apoptotic cells（ P0.05）. This study demonstrated that specific siRNAs targeting Dnmt1 effectively knocked down Dnmt1 mRNA expression in FBFCs,suggesting that the improved SCNT efficacy observed in the siRNA-treated donor cells might be partially due to changes in the cell cycle distribution.

关 键 词：凋亡 细胞周期 供体细胞 增殖 RNA干扰 

分 类 号：Q253[生物学—细胞生物学] Q28