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目的 研究反义脱氧寡核苷酸 (AODN)对培养C6 细胞合成血管内皮生长因子(VEGF)和自身增殖的影响。方法 将不同浓度 (1 .0、2 .5、5 .0、1 0 .0 μmol/L)经硫代修饰的正义、反义、错义寡核苷酸分别加入培养的C6 细胞中 ,采用免疫组织化学SP法检测C6 细胞中VEGF蛋白的表达 ,并用噻唑蓝 (MTT)法检测肿瘤细胞的增殖状况。结果 反义寡核苷酸可抑制C6 细胞VEGF蛋白的表达 ,其抑制作用随AODN浓度的升高而增强 ,错义和正义寡核苷酸却无此作用。其中 ,对照组和浓度为 1 .0、2 .5、5 .0、1 0 .0 μmol/LAODN的VEGF阳性细胞率分别为 82 .1 0 %、73 .1 0 %、70 .0 0 %、57.85 %和 53 .2 0 %。但 3种寡核苷酸对C6 细胞增殖均不发生影响。结论 VEGF AODN药物能抑制VEGF蛋白的产生和分泌 ,可作为胶质瘤基因治疗的新途径
Objective To investigate the effect of antisense oligodeoxynucleotide (AODN) on the proliferation and proliferation of cultured C6 cells induced by vascular endothelial growth factor (VEGF). Methods Sense, antisense and missense oligonucleotides modified with different concentrations (1.0, 2.5, 5.0 and 1.0 μmol / L) were added to cultured C6 cells respectively. The expression of VEGF protein in C6 cells was detected by immunohistochemical SP method. The proliferation of tumor cells was detected by MTT assay. Results Antisense oligodeoxynucleotides inhibited the expression of VEGF protein in C6 cells. The inhibitory effect was enhanced with the increase of AODN concentration, while no effect was observed in the sense and sense oligonucleotides. Among them, the rates of VEGF-positive cells in the control group and the concentrations of 1.0, 2.5, 5.0, and 1.0 μmol / LAODN were 82.1%, 73.1%, 70.0% , 57.85% and 53.2%. However, the three kinds of oligonucleotides did not affect the proliferation of C6 cells. Conclusions VEGF AODN can inhibit the production and secretion of VEGF protein and can be used as a new pathway for gene therapy of glioma