反义寡核苷酸对血管瘤内皮细胞VEGF表达的影响

Effect of Antisense Oligonucleotide on Expression of Vascular Endothelial Growth Factor in Cultured Skin Hemangioma Endothelial Cells

目的探讨反义寡核苷酸(AS-ODN)对培养的血管瘤内皮细胞中血管内皮生长因子(vascular endothelialgrowth factor,VEGF)表达的影响。方法将VEGF反义寡核苷酸(AS-ODN)、正义寡核苷酸(S-ODN)和错义寡核苷酸(M-ODN)分别转染至体外培养的皮肤血管瘤内皮细胞中,采用RT-PCR技术检测各组细胞中VEGF mRNA的表达水平;ELISA法检测各组细胞培养上清液中VEGF蛋白分泌的改变。结果 RT-PCR结果显示:AS-ODN组VEGF mR-NA表达水平较空白对照组明显下降,而S-ODN组、M-ODN组与空白对照组比较,其VEGF mRNA表达未见明显改变。ELISA结果显示:AS-ODN组细胞培养上清液中的VEGF蛋白分泌浓度较空白对照组明显下降,而S-ODN组、M-ODN组与空白对照组比较,VEGF浓度未见明显变化。结论 VEGF AS-ODN转染可以有效地抑制血管瘤内皮细胞VEGF mRNA表达,降低VEGF蛋白分泌,从而有效地抑制血管瘤内皮细胞的生长。

Objective To investigate the impact of antisense oligonucleotides（AS-ODN）on expression of vascular endothelial growth factor（VEGF）in cultured skin hemangioma endothelial cells.Methods AS-ODN,S-ODN and M-ODN were transfected into skin hemangioma endothelial cells respectively.The expression of VEGF mRNA was detected by using RT-PCR.The secretion change of VEGF protein was detected by using ELISA in culture supernatants of skin hemangioma endothelial cells.Results RT-PCR revealed that the expression level of VEGF mRNA in AS-ODN-treated group was significantly decreased as compared with that in blank control group.There was no significant change in the expression of VEGF mRNA in S-ODN-treated group,M-ODN-treated group and control group.ELISA demonstrated that the secretion of VEGF protein in AS-ODN-treated group was significantly decreased as compared with that in control group,but no obvious changes in the secretion of VEGF were found in S-ODN-treated group,M-ODN-treated group and control group.Conclusion VEGF AS-ODN transfection can effectively inhibit the expression of VEGF mRNA in hemangioma vascular endothelial cells and reduce VEGF protein secretion,which can effectively inhibit the growth of hemangioma vascular endothelial cells.