RNA干扰her-2基因对骨肉瘤细胞血管内皮生长因子-A和白细胞介素-8表达的影响  

Effect of silencing her-2 gene by shRNA on expressions of vascular endothelial growth factor-A and interleukin-8 in osteosarcoma cells

作　　者：陈潇[1] 范伟[1] 蔡枭[1] 熊科[1] 胡廷毅[1] 

机构地区：[1]重庆医科大学附属第一医院骨科,重庆400016

出　　处：《中国生物制品学杂志》2012年第11期1459-1463,共5页Chinese Journal of Biologicals

摘　　要：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 silencing her-2 gene by shRNA on the expressions of vascular endothelial growth factor（VEGF）-A and interleukin-8（IL-8） in human osteosarcoma saos-2 cells.Methods Recombinant expression vector with her-2 shRNA was constructed and transfected to saos-2 cells,using the cells untransfected as blank control and those transfected with plasmid shNC as negative control.The transcription levels of her-2,VEGF-A and IL-8 mRNAs in the transfected cells were determined by RT-PCR,while the expression levels of her-2 protein by Western blot.The secretion levels of VEGF-A and IL-8 in cell culture were determined by ELISA.Results The constructed Her-2 shRNA expression plasmid inhibited the expression of her-2 gene,of which the inhibiting rates to her-2 mRNA transcription and her-2 protein expression were 63.05% and 62.59% respectively.Both the mRNA transcription and protein expression levels of VEGF-A and IL-8 in saos-2 cells transfected with her-2 shRNA decreased significantly（P 0.01）.Conclusion Both the mRNA transcription and protein expression levels of VEGF-A and IL-8 decreased significantly after her-2 gene silence,indicating that her-2 gene was involved in the regulation of VEGF-A and IL-8 expressions in osteosarcoma cells and might be used as a novel target for study on the molecular mechanism of angiogenesis of osteosarcoma.

关 键 词：骨肉瘤 HER-2基因 血管内皮生长因子A 白细胞介素8 RNA干扰 

分 类 号：R738.1[医药卫生—肿瘤]