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目的探讨her-2基因沉默对人骨肉瘤细胞株saos-2中血管内皮生长因子-A(Vascular endothelial growth factor-A,VEGF-A)和白细胞介素-8(Interleukin-8,IL-8)表达的影响。方法构建her-2 shRNA重组表达质粒,转染至骨肉瘤细胞株saos-2,同时设空白对照组和shNC阴性对照组;RT-PCR检测her-2、VEGF-A和IL-8基因mRNA的转录水平;Western blot检测her-2蛋白表达的变化;ELISA法检测细胞培养液中VEGF-A和IL-8的分泌水平。结果构建的her-2 shRNA表达载体能抑制her-2基因的表达,对her-2基因mRNA的转录和蛋白表达的抑制率分别为63.05%和62.59%;转染重组质粒her-2 shRNA后VEGF-A和IL-8基因mRNA的转录水平及蛋白分泌水平均显著降低(P<0.01)。结论 her-2基因沉默后,VEGF-A和IL-8基因mRNA的转录水平和蛋白表达水平明显降低,her-2基因参与了骨肉瘤细胞VEGF-A和IL-8基因的表达调控,提示her-2基因可作为研究骨肉瘤血管生成分子机理的新靶点。
Objective To investigate the effect of her-2 gene silencing on the expression of vascular endothelial growth factor-A (VEGF-A) and interleukin-8 (IL-8) in human osteosarcoma cell line saos-2 Impact. Methods The her-2 shRNA recombinant plasmid was constructed and transfected into osteosarcoma cell line saos-2. The blank control group and shNC negative control group were also established. The mRNA expressions of her-2, VEGF-A and IL-8 The level of VEGF-A and IL-8 in the cell culture medium was detected by ELISA. Results The her-2 shRNA expression vector could inhibit the expression of her-2 gene and the inhibitory rates of her-2 gene transcription and protein expression were 63.05% and 62.59%, respectively. After transfected with recombinant plasmid her-2 shRNA, VEGF The mRNA and protein levels of -A and IL-8 mRNA were significantly decreased (P <0.01). Conclusion After her-2 gene silencing, the transcriptional and protein expression of VEGF-A and IL-8 mRNA and protein were significantly decreased. Her-2 gene was involved in the regulation of VEGF-A and IL-8 gene expression in her osteosarcoma cells, -2 gene can be used as a new target for studying the molecular mechanism of osteosarcoma angiogenesis.