文档介绍：PEG10基因siRNA真核表达载体的构建及其对肝癌HepG2细胞凋亡的影响
【关键词】 PEG10基因;RNA干扰;表达载体
【Abstract】 AIM: To construct the siRNA eukaryotic expression vectors of PEG10 gene and to investigate their apoptosisinducing effect on transfected HepG2 cells in vitro. METHODS: Three specific siRNAs of PEG10 were designed, and cloned into the psiRNAhH1neo, which were transiently transfected into HepG2 via Lipofectamine 2000. Realtime quantitative PCR was used to measure the PEG10 mRNA expression in HepG2. The cell growth was assessed by MTT assay and the apoptosis was evaluated by flow cytometry and confocal microscopy. RESULTS: Three specific siRNA eukaryotic expression vectors (siRNA1, siRNA2, siRNA3) targeting PEG10 were essfully produced, which could inhibit the expression of PEG10 mRNA to varying degrees. The expression of PEG10 mRNA was remarkably inhibited at 48 h after transfection, and siRNA2 resulted in the highest inhibition rate (%). HepG2 growth was also inhibited significantly (P<), and cell apoptosis rate reached the highest after siRNA2 transfection (%). CONCLUSION: SiRNA eukaryotic expression vectors targeting PEG10 gene can induce HepG2 cell growth inhibition and apoptosis through downregulating PEG10.
【Keywords】 PEG10 gene; RNA interference; eukaryotic expression vector
【摘要】目的:构建针对遗传印记基因PEG10的siRNA真核表达载体,体外观察其对肝癌HepG2细胞凋亡的影响. 方法:设计针对PEG10基因的3个siRNA靶序列,构建真核表达载体siRNA1,s