Erratum to:J Huazhong Univ Sci Technol[Med Sci]36(4):548–553,2016 https://doi.org/10.1007/s11596-016-1623-6 In the originally published article(https://doi.org/10.1007/s11596-016-1623-6),the immunofluorescence images...Erratum to:J Huazhong Univ Sci Technol[Med Sci]36(4):548–553,2016 https://doi.org/10.1007/s11596-016-1623-6 In the originally published article(https://doi.org/10.1007/s11596-016-1623-6),the immunofluorescence images in shRNA group in Fig.3 were accidentally used rather than the final,formal experiments.To retain consistency,the entire Fig.3 is replaced here with original images of the experiments.The authors declare that this correction will not affect the conclusion of the study.展开更多
[目的]利用shRNA慢病毒载体构建敲低CTNNB1的A549细胞系,检测其对细胞增殖迁移及对趋化因子CCL4表达的影响。[方法]设计靶向CTNNB1基因的shRNA序列,克隆至pLKO.1慢病毒载体中得到重组干扰质粒。运用三质粒系统包装慢病毒;转导A549细胞...[目的]利用shRNA慢病毒载体构建敲低CTNNB1的A549细胞系,检测其对细胞增殖迁移及对趋化因子CCL4表达的影响。[方法]设计靶向CTNNB1基因的shRNA序列,克隆至pLKO.1慢病毒载体中得到重组干扰质粒。运用三质粒系统包装慢病毒;转导A549细胞后用梯度稀释法得到阳性单克隆细胞;采用Western blotting和qRT-PCR检测CTNNB1及其下游基因的mRNA和蛋白质表达水平;采用实时无标记细胞分析实验检测细胞增殖能力,通过细胞划痕实验评估细胞迁移能力。[结果]成功构建稳定敲低CTNNB1的A549细胞系,与A549-shNC相比,A549-shCTNNB1细胞中CTNNB1的mRNA表达降低(1.01±0.05 vs 0.40±0.004,P<0.0001),β-catenin的蛋白表达降低(0.95±0.10 vs 0.47±0.16,P<0.05);细胞增殖有减缓的趋势,增殖相关基因COX2、CCND1、c-MYC的mRNA表达降低(0.978±0.02 vs 0.66±0.09,P<0.01);MMP2/9的mRNA表达降低(0.98±0.001 vs 0.20±0.08,P<0.0001),MMP2/9蛋白表达降低(0.96±0.05 vs 0.70±0.05,P<0.05),细胞迁移能力减弱(0.91±0.02 vs 0.69±0.05,P<0.01);CCL4的mRNA表达上调(1.87±0.39 vs 6.27±0.60,P<0.001)。[结论]敲低CTNNB1的A549细胞增殖速度减缓,迁移能力减弱,且趋化因子CCL4的mRNA表达上调。展开更多
文摘Erratum to:J Huazhong Univ Sci Technol[Med Sci]36(4):548–553,2016 https://doi.org/10.1007/s11596-016-1623-6 In the originally published article(https://doi.org/10.1007/s11596-016-1623-6),the immunofluorescence images in shRNA group in Fig.3 were accidentally used rather than the final,formal experiments.To retain consistency,the entire Fig.3 is replaced here with original images of the experiments.The authors declare that this correction will not affect the conclusion of the study.
文摘[目的]利用shRNA慢病毒载体构建敲低CTNNB1的A549细胞系,检测其对细胞增殖迁移及对趋化因子CCL4表达的影响。[方法]设计靶向CTNNB1基因的shRNA序列,克隆至pLKO.1慢病毒载体中得到重组干扰质粒。运用三质粒系统包装慢病毒;转导A549细胞后用梯度稀释法得到阳性单克隆细胞;采用Western blotting和qRT-PCR检测CTNNB1及其下游基因的mRNA和蛋白质表达水平;采用实时无标记细胞分析实验检测细胞增殖能力,通过细胞划痕实验评估细胞迁移能力。[结果]成功构建稳定敲低CTNNB1的A549细胞系,与A549-shNC相比,A549-shCTNNB1细胞中CTNNB1的mRNA表达降低(1.01±0.05 vs 0.40±0.004,P<0.0001),β-catenin的蛋白表达降低(0.95±0.10 vs 0.47±0.16,P<0.05);细胞增殖有减缓的趋势,增殖相关基因COX2、CCND1、c-MYC的mRNA表达降低(0.978±0.02 vs 0.66±0.09,P<0.01);MMP2/9的mRNA表达降低(0.98±0.001 vs 0.20±0.08,P<0.0001),MMP2/9蛋白表达降低(0.96±0.05 vs 0.70±0.05,P<0.05),细胞迁移能力减弱(0.91±0.02 vs 0.69±0.05,P<0.01);CCL4的mRNA表达上调(1.87±0.39 vs 6.27±0.60,P<0.001)。[结论]敲低CTNNB1的A549细胞增殖速度减缓,迁移能力减弱,且趋化因子CCL4的mRNA表达上调。
