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DLL4/Notch信号途径对A549细胞VEGF表达的影响

Influence of DLL4/Notch signal pathway on the expression of VEGF in A549 cells
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摘要 目的研究Delta-like ligand 4(DLL4)与血管内皮生长因子(VEGF)在人肺腺癌A549细胞中的表达,以及DLL4对VEGF表达的影响。方法 (1)用免疫组化检测DLL4与VEGF在A549细胞中的表达;(2)设计和化学合成靶向DLL4基因的小干扰RNA(siRNA)序列,用lipofectamineTM2000介导DLL4 siRNA转染人A549细胞;(3)提取细胞总RNA,进行反转录-聚合酶链反应(RT-PCR)扩增检测DLL4、VEGF mRNA;(4)用酶联免疫吸附试验(ELISA)检测A549细胞培养上清液中VEGF质量浓度。结果 (1)DLL4、VEGF表达于A549细胞的细胞质内;(2)DLL4 siRNA可使DLL4沉默,同时,明显上调VEGF-A、VEGF-B、VEGF-C mRNA表达,差异有统计学意义(P<0.05);(3)DLL4转染后,细胞培养上清液中VEGF质量浓度明显增加,差异有统计学意义(P<0.01)。结论阻断A549细胞中Notch通路可刺激VEGF过表达。 Objective To study the expression of Delta-like ligand 4 （ DLL4 ） and vascular endothelial growth factor （VEGF） in A549 cells of human lung adenocarcinoma,and the influence of DLL4 on the expression of VEGF. Methods （1） The immunohistochemical method was adopted to detect the expression of DLL4 and VEGF in A549 cells;（2）the small interferon RNA（siRNA） squences of targeted DLL4 gene were designed and chemically synthesized,and the DLL4 siRNAs were transfect-ed into A549 cells by lipofectamineTM2000;（3）total RNA was extracted from A549 cells to conduct reversal transcription poly merase chain reaction（RT-PCR） for amplifying and detecting DLL4 and VEGF mRNA;（4）the enzyme linked immunosorbent assay（ELISA） were applied to determine the mass concentration of VEGF in culture supernatant fluid of A549 cells. Results （1）DLL4 and VEGF were expressed in cytoplasm of A549 cells;（2）the siRAN of DLL4 gene could make DLL4 gene silence,and up-regulate the expression of VEGF-A mRNA,VEGF-B mRNA amd VEGF-C mRNA with statistically significant difference （P＆lt;0.05）;（3）after transfection of DLL4 cells,the mass concentration of VEGF in culture supernatant fluid increased obviously with statistically sig-nificant difference（P＆lt;0.01）. Conclusion Blocking Notch signal pathway in A549 cells can stimulate the expression of VEGF.
作者 孙晓东 王珺 王燕 田宗文
机构地区 武汉大学医学院人体解剖学与组织胚胎学系 湖北医药学院基础医学研究所
出处 《现代医药卫生》 2014年第7期963-965,共3页 Journal of Modern Medicine & Health
基金 湖北省十堰市科技项目(十科发2010-047S 十科通[2006]18K33)
关键词 酶联免疫吸附测定 RNA干扰 DLL4 NOTCH信号途径 A549细胞 血管内皮生长因子A Vascular endothelial growth factor A Enzyme-linked immunosorbent assay RNA interference DLL4/notch signal pathway A549 cells
分类号 R734.2 [医药卫生—肿瘤]
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参考文献11

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二级参考文献40

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现代医药卫生
2014年 第7期

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