载体携带小干扰RNA抑制Tca8113细胞血管内皮生长因子的表达  

Suppression of vascular endothelial growth factor expression by vector-based small interfering RNA in human tongue squamous carcinoma cell line Tca8113

作　　者：于大海[1] 曹莹[1] 姚志文[1] 李敬[1] 陈海波[1] 郝洁[1] 

机构地区：[1]广西医科大学附属口腔医院口腔颌面外科,广西南宁530021

出　　处：《华西口腔医学杂志》2008年第5期550-552,共3页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(30360111)

摘　　要：目的了解小干扰RNA(siRNA)对人舌癌细胞株Tca8113血管内皮生长因子(VEGF)表达的抑制作用。方法构建2对针对血管内皮生长因子的siRNA真核表达载体(Pu-VEGF-siRNA1,Pu-VEGF-siRNA2),经脂质体Lipo-fectamine 2000转染Tca8113细胞,以空载体组作实验对照组,未转染组作空白对照组。采用逆转录聚合酶链反应(RT-PCR)、免疫组化、酶联免疫吸附(ELISA)方法分别检测转染后的Tca8113细胞VEGF mRNA及蛋白表达。结果与实验对照组和空白对照相比较,转染Pu-VEGF-siRNA1和Pu-VEGF-siRNA2后Tca8113细胞的VEGF mRNA和蛋白表达明显下降。结论通过RNA干扰技术能有效抑制舌癌Tca8113细胞VEGF mRNA和蛋白的表达。Objective To assess suppression effects of vector-based small interfering RNA （siRNA） on vascular endothelial growth factor （VEGF） expression of human tongue squamous carcinoma cell line （Tca8113） in vitro. Methods Two siRNA targeting VEGF constructed in eukaryotic expression vector（Pu-VEGF-siRNA1, Pu-VEGF- siRNA2）, eukaryotic expression vector as the experiment control, all of which were transfected into Tca8113 cells with Lipofectamine 2000. Non-transfection cell was used as negative control. VEGF mRNA and protein were detected by reverse transcription polymerase chain reaction（RT-PCR）, immunohistochemistry and enzyme linked immunosorbent assay （ELISA）, respectively. Results Compared to the experimental and negative controls, the expression of VEGF mRNA and protein were significantly decreased in the Pu-VEGF-siRNA1 group and Pu-VEGF-siRNA2 group. But there were no significant differences between two controls （P〉0.05）. Conclusion Vector-based siRNAs targeting VEGF are efficient in down-regulating VEGF expression in Tca8113 cells.

关 键 词：舌癌 血管内皮生长因子 小干扰RNA 

分 类 号：R739.8[医药卫生—肿瘤]