CD、VEGFR-1基因对恶性胶质瘤细胞和血管内皮细胞共培养体系中VEGF表达及细胞增殖的影响  

Effects of CD and VEGFR-1 genes on VEGF expression and cell proliferation in the co-culture of malignant glioma cells and vascular endothelial cells

作　　者：李振业[1] 张鹏飞[1] 周建华[2] 周婉琦 王红云[1] LI Zhen-ye;ZHANG Peng-fei;ZHOU Jian-hua(Department of Neurosurgery,Affiliated TianTan Hospital,Capital University of Medical Sciences,Beijing Neurosurgical Institue,Beijing 100070,China;Institute of Materia Medica,Chinese Academy of Medical Sciences,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京天坛医院神经外科北京市神经外科研究所,北京100070 [2]中国医学科学院药物研究所,北京100050

出　　处：《临床和实验医学杂志》2018年第24期2617-2620,共4页Journal of Clinical and Experimental Medicine

基　　金：北京市自然科学基金资助项目(编号:7072015);北京市科技新星计划资助项目(编号:2007A042)

摘　　要：目的研究胞嘧啶脱氨酶(CD)、血管内皮生长因子受体1(VEGFR-1)基因共表达质粒对恶性胶质瘤细胞和血管内皮细胞共培养体系血管内皮生长因子(VEGF)表达及细胞增殖的抑制作用。方法转染组:CD基因、VEGFR-1基因共表达质粒p EGFP-C1-CD-VEGFR-1转染人血管内皮细胞共培养体系中的人恶性胶质瘤细胞,未转染组:未做细胞转染,对照组:转染空载体。酶联免疫吸附测定(ELISA)法检测培养液上清中VEGF含量,原位杂交及免疫组化检测细胞VEGFmRNA及VEGF的表达,通过流式细胞术观察其对细胞活性的影响。结果转染后转染组细胞VEGFmRNA为0. 62±0. 06,VEGF为0. 42±0. 04,较未转染组及对照组显著降低,培养液中VEGF含量为60. 34±31. 22,较未转染组及对照组显著降低,细胞活性受到抑制,流式细胞术发现转染组细胞的G1期细胞比率明显高于未转染组及对照组,细胞出现凋亡。结论 CD、VEGFR-1基因共表达质粒可抑制恶性胶质瘤细胞和血管内皮细胞共培养体系中VEGFmRNA及VEGF的表达,抑制细胞活性、诱导细胞凋亡。Objective To study the inhibitory effect of CD,VEGFR-1 gene coexpression plasmid on VEGF expression and cell proliferation in malignant glioma cells and vascular endothelial cell co-culture system.Methods Transfection group:cytosine deaminase(CD)gene,vascular endothelial growth factor receptor 1(VEGFR-1)gene co-expression plasmid pEGFP-C1-CDVEGFR-1 transfection Human vascular endothelial cell co-culture human malignant glioma cells in the system.Untransfected group:not transfected cells.Control group:transfected empty carrier.Enzyme-linked immunoadsorption assay(ELISA)was used to detect the content of vascular endothelial growth factor(VEGF)in culture solution,and in situ hybridization and immunohistochemistry were used to detect the expression of VEGFmRNA and VEGF cells.Flow cytometry was used to observe cell activity.Results After transfection,VEGFmRNA in the transfection group was 0.62±0.06,VEGF was 0.42±0.04,which was significantly lower than that in the untransfected group and the control group.The content of VEGF in the culture was 60.34±31.22,which was significantly lower than that in the untransfected group and the control group.Cell activity was inhibited.Flow cytometry showed that the ratio of G1 cells in transfection group was significantly higher than that in untransfected group and control group,and apoptosis occurred.Conclusion The coexpression of plasmids in CD and VEGFR-1 genes can inhibit the expression of VEGFmRNA and VEGF in the co-culture system of malignant glioma cells and vascular endothelial cells,inhibit cell activity,and induce apoptosis.

关 键 词：CD基因 VEGFR-1基因 恶性胶质瘤 血管内皮细胞 

分 类 号：R739.4[医药卫生—肿瘤]