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论文摘要

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 曹莹 姚志文 李敬 陈海波 郝洁

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

收稿日期:2008-10-25          年卷(期)页码:2008,26(05):550-

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:舌癌,血管内皮生长因子,小干扰RNA,

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

基金项目:

国家自然科学基金资助项目(30360111)

中文摘要

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。

英文摘要

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.

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