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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 To study the inhibitory effect of CD, Flt-1 gene on VEGF expression and growth in malignant glioma cells. Methods The malignant glioma cells were co-transfected with CD, FLT-1 gene plasmid pEGFP-C1-CD-Flt-1. VEGF levels in the supernatant of the culture supernatants were detected by ELISA. The expression of VEGF mRNA and protein The cell viability was observed by MTT assay and flow cytometry. Results The mRNA and protein levels of VEGF in the experimental group decreased significantly after 72h, and the VEGF content in the culture medium decreased. The cell viability was inhibited by flow cytometry. The percentage of cells in the G1 phase in the experimental group was significantly higher than that in the control group . Conclusion The recombinant plasmid pEGFPC1-CD-Flt-1 can inhibit the expression of VEGF mRNA and protein in malignant glioma cells in vitro and inhibit cell viability and induce apoptosis.