Impact of Co-transfection with Livin and Survivin shRNA Expression Vectors on Biological Behavior of HepG2 Cells

作者：Wei Xu, Chang Hong, Chengkun Qin, Yunpeng Zhai · 发表于：Asian Pacific Journal of Cancer Prevention · 年份：2013 · DOI：10.7314/apjcp.2013.14.9.5467 · 被引用次数：5 · 研究领域：RNA Interference and Gene Delivery、Cell death mechanisms and regulation、CRISPR and Genetic Engineering

OBJECTIVE: To construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting Livin and Survivin genes, and to explore the impact of co-transfection of Livin and Survivin shRNA expression vectors on the biological behavior of HepG2 cells. METHODS: shRNA eukaryotic expression vectors pSD11-Livin and pSD11- Survivin were designed and constructed then transfected into HepG2 cells separately or in combination. mRNA and protein expression in transfected cells was assessed by quantitative fluorescence PCR and Western blotting, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by TUNEL assay. RESULTS: The Livin and Survivin shRNA eukaryotic expression vectors were successfully constructed and transfected into HepG2 cells. The relative mRNA expression levels of Livin and Survivin in HepG2 cells co-transfected with pSD11-Livin and pSD11-Survivin were 0.12 ± 0.02 and 0.33 ± 0.13, respectively, which was significantly lower than levels in cells transfected with either pSD11-Livin or pSD11-Survivin (P<0.05). The relative protein expression levels of Livin and Survivin in the co-transfected cells were also significantly decreased compared to single- transfection (P<0.05). The inhibition rate of cell growth in the co-transfection group was higher than that in the single-transfection groups at 48 h, 60 h, or 72 h after transfection (P<0.01). The apoptotic rate increased to the greatest extent in the co-transfection group relative to any other group (P...