反义脱氧寡核苷酸对培养C_6胶质瘤细胞合成血管内皮生长因子和细胞增殖的影响

Effects of antisense oligodeoxynucleotides on vascular endothelial growth factor expression and cell proliferation in C_6 glioma in vitro experiment

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 。

Objective To investigate the effects of antisense oligodeoxynucleotides on VEGF expression and C 6 glioma cell proliferation.Methods The synthesized antisense,missense and sense oligodeoxynucleotides at different concentrations (1.0,2.5,5.0,10.0?μmol/L) were added to the medium of cultured C 6 glioma cells.VEGF protein expression was detected by SP immuno histochemical technique.Cell proliferation was assessed using MTT method.Results Compared with the control,VEGF protein expression was inhibited by antisense oligodeoxynucleotides.And the inhibitory effects were increased with the increasing concentration.However,no inhibitory effect was found in the cell lines after addition of missense and sense oligodeoxynucleotides.Neither antisense oligodeoxynucleotides nor missense and sense oligodeoxynucleotides had effect on tumor cell proliferation in vitro experiment.Conclusion VEGF antisense oligodeoxynucleotides can be used as a new gene therapy for glioma through its antiangiogenesis effect by inhibition of VEGF.