华西口腔医学杂志 ›› 2008, Vol. 26 ›› Issue (05): 550-552.

• 专栏论著 • 上一篇    下一篇

载体携带小干扰RNA抑制Tca8113细胞血管内皮生长因子的表达

于大海 曹莹 姚志文 李敬 陈海波 郝洁   

  1. 广西医科大学附属口腔医院口腔颌面外科, 广西南宁530021
  • 收稿日期:2008-10-25 修回日期:2008-10-25 出版日期:2008-10-20 发布日期:2008-10-20
  • 通讯作者: 于大海,Tel:0771-3277213
  • 作者简介:于大海(1968-),男,广西人,教授,博士
  • 基金资助:

    国家自然科学基金资助项目(30360111)

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

YU Da-hai, CAO Ying, YAO Zhi-wen, LI Jing, CHEN Hai-bo, HAO Jie   

  1. Dept. of Oral and Maxillofacial Surgery, College of Stomatology, Guangxi Medical University, Nanning 530021, China
  • Received:2008-10-25 Revised:2008-10-25 Online:2008-10-20 Published:2008-10-20
  • Contact: YU Da-hai,Tel:0771-3277213

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Lipofectamine2000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。

关键词: 舌癌, 血管内皮生长因子, 小干扰RNA

Abstract:

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-VEGFsiRNA2),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.

Key words: tongue cancer, vascular endothelial growth factor, small interfering RNA