靶向核因子κB p65siRNA促人血管内皮细胞EAhy926的凋亡

Targeting nuclear factor-kappa B p65siRNA promotes apoptosis of human vascular endothelial cell EAhy926

背景:作者前期工作已经证实,子宫内膜异位症患者的在位、异位内膜中核因子κBmRNA和蛋白均表达升高,核因子κB参与了子宫内膜异位症细胞黏附、侵袭和血管形成的病理过程。目的:观察RNA干扰技术(RNAi)靶向抑制核因子κB p65基因对人血管内皮细胞(EAhy926)增殖的影响。方法:10μg/L血管内皮生长因子刺激下培养EAhy926细胞,给予转染核因子κB p65 siRNA,同时设立对照组,分别是阴性对照siRNA转染组、空白脂质体组及正常对照组(不给予其他任何处理),48h后分别采用Heochst 33342荧光检测法、MTT法及流式细胞仪术检测细胞的增殖抑制与凋亡情况。结果与结论:与正常对照组比较,核因子κB p65 siRNA组的细胞增殖抑制率及细胞凋亡率均明显升高(P<0.05);而阴性对照siRNA转染组及空白脂质体组的细胞增殖抑制率及细胞凋亡率则差异无显著意义。结果证实,核因子κB p65 siRNA能显著抑制人血管内皮细胞增殖,促进内皮细胞的凋亡发生。

BACKGROUND：Previous research has demonstrated that nuclear factor（NF） κB mRNA and protein are both increased in eutopic and ectopic endometrium of patients with endometriosis.NF κB participates in the pathological process of cell adhesion,invasion and angiopoiesis in endometriosis.OBJECTIVE：To investigate the effect of small interfering RNA（siRNA） targeting NF-ΚB on proliferation of human vascular endothelial cells（EAhy926 cells）.METHODS：EAhy926 cells were cultured with stimulation of vascular endothelial growth factor（VEGF,10 μg/L）,and then were transfected with NF-ΚB-siRNA.Meanwhile,negative control-siRNA,empty liposome and normal control groups were established.The proliferation inhibition and apoptosis of cells were detected by Heochst 33342 test,MMT assay and flow cytometry analysis.RESULTS AND CONCLUSION：Compared with the normal control group,the rates of proliferation inhibition and apoptosis of cells were increased obviously in the NF-ΚB p65-siRNA group（P 0.05）;there were no significant differences between empty liposome group,negative control siRNA group and normal control group,respectively（P 0.05）.Synthetic siRNA can effectively inhibit the proliferation and stimulates the apoptosis of human vascular endotheliar cells.