CASK在血管内皮细胞缺氧应激增殖反应中的作用  

Role of calcium/calmodulin-dependent serine protein kinase in hypoixa related endothelial cell proliferation

作　　者：苏踊跃[1] 孙林辉[1] 邬江[1] 徐永清[1] 陆声[1] 陈兵[1] 罗向东[2] 许波[3] 

机构地区：[1]成都军区昆明总医院骨科,昆明650032 [2]第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆400038 [3]第三军医大学西南医院口腔科,重庆400038

出　　处：《第三军医大学学报》2010年第14期1546-1548,共3页Journal of Third Military Medical University

摘　　要：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 screen an endothelial cell line with calcium/calmodulin-dependent serine protein kinase （CASK） knockdown and explore the cells’ reaction to hypoxia stress. Methods CASK RNA interference plasmid pGenesil-siCASK and control plasmid pGenesil-Negative were generated,and then transfected into EA.hy926 endothelial cells with liposome followed by screened with G418. The expression of CASK was detected in normal,pGenesil-siCASK and pGenesil-Negative transfected cells with Western blotting. Hypoxic stress was induced in the transfected and untransfected cells cultured under an atmosphere with an airflow （94%N2,5%CO2 and 1%O2） at 10 L/min in an interval of 5 min inflow and 5 min stillness for 3 times. After the cells were cultured for 0,1,3,6 and 12 h,the cell cycle was investigated by using flow cytometric analysis. Results Western blotting results showed that CASK was mildly expressed in the EA.hy926 cells after pGenesil-siCASK transfection,while its expression had no obvious change in those transfected with pGenesil-negative. The cell proliferation indexes of EA.hy926 cells were elevated by short term hypoxia （0 h： 42.93%,1 h： 50.46% and 3 h： 52.53%）,while were decreased with the prolongation of hypoxic duration （6 h： 40.71%,12 h： 25.18%）. However,the proliferation indexes of CASK-knockdown endothelial cells were not changed significantly under transient hypoxic stress. Conclusion CASK knockdown endothelial cells have less proliferation changes than EA.hy926 endothelial cells under short term hypoxic stress.

关 键 词：缺氧 钙/钙调蛋白依赖性丝氨酸蛋白激酶 血管内皮细胞 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R345.57[医药卫生—基础医学]