| 可溶性血管内皮生长因子受体1真核克隆表达及其对血管内皮细胞增殖的影响 |
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| 引用本文： | 姬玲玲,毛淑华,刘洪,许士叶,杨雨,易成,黄英. 可溶性血管内皮生长因子受体1真核克隆表达及其对血管内皮细胞增殖的影响[J]. 生物医学工程学杂志, 2010, 0(2) |
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| 作者姓名： | 姬玲玲 毛淑华 刘洪 许士叶 杨雨 易成 黄英 |
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| 作者单位： | 四川大学华西基础医学与法医学院病理生理学教研室;成都医学院门诊部;四川大学华西医院腹部肿瘤科; |
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| 基金项目： | 国家自然科学基金资助项目(30570693); 四川省科技厅攻关计划科研基金资助项目(2006Z09-030) |
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| 摘 要： | 本研究旨在构建可溶性血管内皮生长因子(vascular endothelial growth factor,VEGF)受体1(soluble fmslike tyosine kinase-1,sFlt-1)的真核表达质粒pcDNA3.1-sFlt-1,并观察sFlt-1对血管内皮细胞增殖的影响。提取人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)总RNA,扩增Flt-1基因胞外1-3结构域,构建真核表达质粒pcDNA3.1-sFlt-1,测序鉴定基因序列。将重组质粒转染Lewis肺癌细胞,采用RT-PCR和SDS-PAGE检测sFlt-1在基因及蛋白水平的表达情况。MTT法检测sFlt-1对VEGF诱导的HUVECs生长的影响。结果显示:①插入片段序列正确;②sFlt-1在基因水平成功表达且转染后的Lewis肺癌细胞能分泌表达sFlt-1;③含sFlt-1的细胞上清液可明显抑制VEGF诱导的HUVECs增殖。本研究成功构建了真核表达质粒pcDNA3.1-sFlt-1,sFlt-1,在基因和蛋白水平均获得有效表达,且表达的蛋白可明显抑制由VEGF诱导...
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| 关 键 词： | 血管内皮生长因子 可溶性血管内皮生长因子受体1 内皮细胞 真核表达 抗血管生成 |
Construction and Expression of Soluble Vascular Endothelial Growth Factor Receptor-1 Eukaryotic Expression Vector and Its Effect on Proliferation of Vascular Endothelial Cells |
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| Ji Lingling Mao Shuhua Liu Hong, Xu Shiye Yang Yu Yi Cheng Huang Ying. Construction and Expression of Soluble Vascular Endothelial Growth Factor Receptor-1 Eukaryotic Expression Vector and Its Effect on Proliferation of Vascular Endothelial Cells[J]. Journal of biomedical engineering, 2010, 0(2) |
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| Authors: | Ji Lingling Mao Shuhua Liu Hong Xu Shiye Yang Yu Yi Cheng Huang Ying |
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| Affiliation: | Ji Lingling1 Mao Shuhua1 Liu Hong1,2 Xu Shiye1 Yang Yu3 Yi Cheng3 Huang Ying1 1(Department of Pathophysiology,West China School of Preclinical , Forensic Medicine,Sichuan University,Chengdu 610041,China)2(Department of Out-patient,Chengdu Medical College,Chengdu 610083,China)3(Division of Abdominal Cancer,West China Hospital,China) |
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| Abstract: | This study sought to construct recombinant eukaryotic plasmid pcDNA3.1-sFlt-1 and observe its effect on proliferation of vascular endothelial cells.Total RNA was extracted from human umbilical vein endothelial cells(HUVECs) firstly.The 1st-3rd Ig-like domains of Flt were amplified by polymerase chain reaction(PCR) from the full-length cDNA.Subsequently,the PCR product was cloned into the eukaryotic plasmid pcDNA3.1(+)/myc-His.The constructed recombinant plasmid pcDNA3.1-sFlt-1 was sequenced.Then recombinant... |
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| Keywords: | Vascular endothelial growth factor(VEGF) Soluble vascular endothelial growth factor receptor-1(sFlt-1) Endothelial cell Eukaryotic expression Anti-angiogenesis |
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