于孟飞,王文璐,易建明.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[J].中南民族大学学报自然科学版,2018,(2):24-30
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
Interference of DNA Methyltransferase 1 Gene Silenced by siRNA on Fetal Bovine Fibroblast Cell Proliferation and Apoptosis
  
DOI:
中文关键词: 凋亡  细胞周期  供体细胞  增殖  RNA干扰
英文关键词: apoptosis  cell cycle  donor cell  proliferation  RNA interference
基金项目:国家自然科学基金青年项目(81400252);中央高校基本科研业务费专项资金资助项目(2013PY130)
作者单位
于孟飞1,2,王文璐2,易建明1,2,* 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 430070 
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中文摘要:
      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.
英文摘要:
      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 (P<0.05). This study demonstrated that specific siRNAs targeting Dnmt1effectively 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.
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