下调血管内皮细胞蛋白激酶CK2表达对血管增殖影响的体外研究

目的 探究蛋白激酶CK2表达下调对血管内皮细胞形成新生血管的能力以及对其细胞中血管内皮生长因子(VEGF)表达的影响。方法 化学合成siRNA转染人脐静脉内皮细胞(HUVECs), 用RT-PCR法和Western blotting法检测细胞转染效率;MTT法检测CK2表达下调对HUVECs细胞增殖的影响;成管实验和划痕实验检测CK2表达下调对细胞成管、迁移的影响;Real-time PCR和Western blotting法检测siRNA干扰蛋白激酶CK2表达后VEGF mRNA及蛋白的表达。结果 siRNA干扰蛋白激酶CK2达到了预期干扰率75%, 蛋白激酶CK2表达下调后抑制了HUVECs增殖、成管、迁移(P<0.05), 也抑制了VEGF mRNA及蛋白的表达。结论 蛋白激酶CK2表达下调可抑制VEGF的表达及HUVECs的增殖、成管、迁移。

In vitro study of the effect of Casein Kinase2 siRNA on anginogesis

Objective To prove the effect of Casein Kinase2 siRNA on vascular endothelial growth factor(VEGF) expression and the angiogenic capacity in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were transfected with either Casein Kinase2 siRNA or negative control siRNA. Cell proliferation was analyzed by MTT assay; the angiogenic capacity was accessed by tube formation; cell migration was assessed by the wound-healing model. Gene and protein expressions of VEGF were examined by Real-time PCR and Western blotting methods. Results Casein Kinase2 siRNA decreased the expressions of Casein Kinase2 and declined HUVEC proliferation, tube formation and migration. It was also reduced the expression of VEGF mRNA and VEGF protein. Conclusion Our results demonstrated that Casein Kinase2 siRNA reduced the expression of VEGF and declined the capacity of angiogenesis in HUVECs.