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目的建立钙/钙调蛋白依赖性丝氨酸蛋白激酶(calcium/calmodulin-dependent serine protein kinase,CASK)剔降血管内皮细胞株,初步探讨其对短暂缺氧的反应性。方法采用RNAi技术和脂质体介导的细胞转染技术,建立CASK表达下调的血管内皮细胞株,并观察缺氧应激条件下培养0(常氧)、1、3、6、12h细胞增殖指数的变化。结果血管内皮细胞在短暂缺氧后细胞增殖指数升高(常氧:42.93%,1h:50.46%,3h:52.53%),随着缺氧时间延长增殖指数则低于常氧对照组(6h:40.71%,12h:25.18%)。而CASK剔降血管内皮细胞株,在1～12h缺氧应激时限内,其增殖指数无明显变化。结论 CASK剔降内皮细胞株与对照EA.hy926细胞比较,对短暂缺氧应激的增殖反应性减弱。
OBJECTIVE: To establish a calcium / calmodulin-dependent serine protein kinase (CASK) knockout vascular endothelial cell line to investigate its reactivity to transient hypoxia. Methods RNAi and lipofectamine mediated cell transfection were used to establish the vascular endothelial cell line with CASK downregulation and to observe the effects of 0 (normoxia), 1, 3, 6, 12h cell proliferation under hypoxia stress Changes in the index. Results The proliferation index of vascular endothelial cells was increased after transient hypoxia (normoxia: 42.93%, 1h: 50.46%, 3h: 52.53% 40.71%, 12h: 25.18%). The CASK knockdown vascular endothelial cell line, in 1 ~ 12h hypoxia stress time limit, the proliferation index no significant change. CONCLUSIONS: CASK knockdown endothelial cell line attenuates the proliferative response to transient hypoxia stress compared with control EA.hy926 cells.