Department of Cardiothoracic Surgery, The Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, 226001, Jiangsu, People's Republic of China.
Mol Cell Biochem. 2014 Jan;386(1-2):117-24. doi: 10.1007/s11010-013-1850-0. Epub 2013 Oct 26.
The aim of this study was to investigate the inhibitory effect of a siRNA cocktail targeting Vascular endothelial growth factor (VEGF) and Human epidermal growth factor receptor 2 (HER2) on cell proliferation, induced apoptosis and the expression of VEGF and HER2 in human gastric carcinoma cell. The silencing rate of pre-designed siRNAs that targeted VEGF and HER2 was detected by Real-time Quantitative PCR (RT-QPCR) analysis. Furthermore, the best silencing siRNA that targeted VEGF and HER2 was prepared as a cocktail to co-knockdown VEGF and HER2 expression at both mRNA and protein levels which were detected by RT-QPCR and Western blot analysis. Cell proliferation inhibition rates were determined by CCK8 assay. The effect of siRNA cocktail on cell apoptosis was determined by flow cytometry. The migration inhibition of siRNA cocktail was analyzed by wound-healing assay. The ability of VEGF to induce endothelial cells to proliferate was examined in HUVECs by the method of tube formation assay. The pre-designed siRNAs could inhibit VEGF and HER2 mRNA level. siRNA cocktail, and co-downregulation of VEGF and HER2 result in significant inhibition of gastric cancer growth and migration in vitro. The inhibition of VEGF and HER2 expressions can induce apoptosis of SGC-7901 cells.
本研究旨在探讨针对血管内皮生长因子（VEGF）和人类表皮生长因子受体 2（HER2）的 siRNA 鸡尾酒对人胃癌细胞增殖、诱导凋亡以及 VEGF 和 HER2 表达的抑制作用。通过实时定量 PCR（RT-QPCR）分析检测针对 VEGF 和 HER2 的预设计 siRNA 的沉默率。此外，制备针对 VEGF 和 HER2 的最佳沉默 siRNA 作为鸡尾酒，共同下调 VEGF 和 HER2 在 mRNA 和蛋白水平的表达，通过 RT-QPCR 和 Western blot 分析进行检测。通过 CCK8 测定法确定细胞增殖抑制率。通过流式细胞术测定 siRNA 鸡尾酒对细胞凋亡的影响。通过划痕愈合试验分析 siRNA 鸡尾酒的迁移抑制作用。通过管形成试验检查 VEGF 诱导血管内皮细胞增殖的能力。预设计的 siRNA 可抑制 VEGF 和 HER2 mRNA 水平。siRNA 鸡尾酒和 VEGF 和 HER2 的共同下调导致体外胃癌生长和迁移的显著抑制。抑制 VEGF 和 HER2 的表达可诱导 SGC-7901 细胞凋亡。
