摘要
目的探讨survivin基因沉默对人结肠癌Lovo细胞体外生长的抑制作用。方法构建靶向survivin的shRNA载体质粒,然后转染结肠癌Lovo细胞。用RT-PCR和Western Blot方法检测survivin的mRNA和蛋白的表达情况。MTT实验评价转染后各组细胞的增殖抑制情况。结果靶向survivin的shRNA能够明显下调survivin mRNA和蛋白质表达(P<0.05),其抑制率分别是39.59%和42.32%。MTT实验提示转染特异性靶向survivin shRNA的Lovo细胞在转染48小时后出现呈时间依赖性的生长抑制。在转染后72h,该细胞存活率仅为41.8%,与空白对照组相比有显著性的降低(P<0.05)。结论靶向survivin的特异性shRNA能够有效地沉默结肠癌Lovo细胞的survivin基因表达,体外实验能显著抑制肿瘤细胞增殖。
Objective To investigate the inhibitory effect of surviving-shRNA on survivin gene expression and cell proliferation of the human colon cancer cell.Methods The survivin-shRNA was constructed and transfected into Lovo cells with Lipofectamine.RT-PCR and western blot analysis were used to detect down-regulations of survivin mRNA and protein expressions respectively.Cell proliferation inhibition was determined using MTT assay.Results The expressions of survivin mRNA and protein were successfully suppressed by survivin-shRNA(P<0.05),with inhibition rates of 39.59% and 42.32% respectively.MTT assay indicated that the specific shRNA inhibited tumor cell growth in a time-dependent manner since 48 hours post-transfection.The cell viability of surviving-shRNA transfected group decreased significantly to 41.8% at 72 h,which was of statistical significance compared with that of blank control group(P<0.05).Conclusion The specific survivin-shRNA could down-regulate the expression of survivin gene and inhibit the growth of Lovo cells in vitro.[
出处
《消化肿瘤杂志（电子版）》
2010年第3期167-170,共4页
Journal of Digestive Oncology(Electronic Version)
基金
广东省科技计划资助项目(编号2010B080701038)
