CD、Flt-1基因对原代培养的恶性胶质瘤细胞VEGF表达及生长的影响

Expression of VEGF and proliferation in maligant glioma primary culturre cells in association with recombinant plasmid coexpressing CD and Flt-1 genes transfection

目的研究CD、Flt-1基因对恶性胶质瘤细胞VEGF表达及生长的抑制作用。方法 CD、FLT-1基因共表达质粒pEGFP-C1-CD-Flt-1转染恶性胶质瘤细胞,ELISA法检测培养液上清中VEGF含量,实时定量PCR及免疫组化检测细胞VEGFmRNA及蛋白的表达,通过MTT法和流式细胞术观察其对细胞活性的影响。结果转染72h后实验组细胞VEGFmRNA及蛋白水平显著降低,培养液中VEGF含量降低,细胞活性受到抑制,流式细胞术发现实验组细胞的G1期细胞比例明显高于对照组,细胞出现凋亡。结论重组质粒pEGFPC1-CD-Flt-1,其体外可抑制恶性胶质瘤细胞VEGFmRNA及蛋白的表达,抑制细胞活性、诱导细胞凋亡。

Objective The study was designed to observe whether recombinant plasmid coexpressing CD and Flt-1genes transfection to maligant glioma primary culturre cells are associated with its expression of VEGF and proliferation.Methods The recombinant plasmid coexpressing CD and Flt-1genes was infected into maligant glioma primary culturre cells,the quantity of VEGF mRNA and protein was detected by in situ hybridization and immunohistochemistry.The quantity of VEGF protein in supernatant was detected by ELISA.Proliferation abilities were measured by MTT and FCM.Results 72 hours after transfection,VEGF mRNA protein levels within the cells and in culture medium both decreased significantly.The proliferation rate of cells was inhibited.Cells numbers of G1 phrase were significantly higher than those in control group.Cell apoptosis was found.Conclusion The recombinant plasmid coexpressing CD and Flt-1genes could inhibit expression of VEGF and the cells proliferation and induce their apoptosis in vitro.