| 摘要: |
| 目的:观察人血管内皮生长因子受体(VEGFR)RNA干扰重组腺病毒(Ad-VEGFRshRNA)对人肺腺癌细胞株A549生长的抑制作用。方法:制备携带人VEGFR的RNA干扰重组腺病毒,转染A549细胞,通过荧光显微镜观察转染效率。应用Western印迹法检测A549细胞VEGFR 蛋白的表达,MTT比色法测定活细胞数并绘制细胞生长曲线,细胞周期及集落形成试验测定细胞生长抑制情况。同时,制备裸鼠A549细胞移植瘤模型,观察肿瘤生长情况。结果:Western印迹法显示RNA干扰组VEGFR表达水平明显降低；细胞生长曲线表明,RNA干扰后细胞生长明显减缓;细胞周期及集落形成试验提示RNA干扰后肿瘤细胞生长受到明显抑制。裸鼠皮下移植瘤生长情况提示RNA干扰可以显著抑制肿瘤生长。结论:RNA干扰重组腺病毒Ad-VEGFRshRNA能有效抑制A549细胞中VEGFR的表达,并能抑制肿瘤生长。 |
| 关键词: RNA干扰 血管内皮生长因子受体 腺病毒 肺腺癌细胞 |
| DOI：10.3724/SP.J.1008.2008.00 |
| 投稿时间：2008-01-31修订日期：2008-04-21 |
| 基金项目:上海市科委重点项目(04JC14006). |
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| Vascular endothelial growth factor receptor targeted RNA interference inhibits growth of human lung adenocarcinoma cells |
| ZHANG Guan-xin1, HOU Xiao-lei2, LI Bai-ling1*,HUANG Sheng-dong1 |
| (1.Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433,China;2.Department of ICU, Yueyang Hospital of Integrative Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437) |
| Abstract: |
| Objective:To investigate the inhibitory effects of RNA silencing via adenovirus-mediated vascular endothelial growth factor receptor (VEGFR) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo．Methods: Ad-VEGFRshRNA adenovirus containing enhanced green fluorescent protein (EGFP) gene and VEGFRshRNA was constructed and was used to infect A549 cells; fluorescent microscopy was used to observe the infection efficiency. Western blotting assay was used to examine the expression of VEGFR protein in A549 cells. MTT method was used to examine the cell viability and the cell growth curve was drawn. The inhibition of cell growth was examined by cell cycle and colony-forming test. Meanwhile, nude mice were transplanted with A549 cells to establish tumor-bearing model, and the long term growth of tumor was observed.Results: Western blotting revealed that the expression of VEGFR was obviously decreased in the RNA interference group. The cell growth curve indicated that the cell growth was obviously inhibited after RNA interference. Cell cycle and colony-forming test indicated that the tumor growth was obviously inhibited after RNA interference. In vivo study with nude mice also indicated that RNA interference obviously inhibited tumor growth.Conclusion: The constructed VEGFR-targeted shRNA can effectively inhibit VEGFR expression in A549 cells and can suppress the growth of A549 cells. |
| Key words: RNA inteference vascular endothelial growth factor receptor adenovirus lung adenocarcinoma cells |
