敲低IK1钾通道抑制ClC-3氯通道的表达和功能  

Knockdown of IK1 potassium channel inhibits expression and function of ClC-3 chloride channel

作　　者：邹丽莉[1] 叶东[2] 吕瑞玲[1] 汪源[3] 孙晓雪[3] 朱林燕[3] 陈丽新[3] 王立伟[1] 

机构地区：[1]暨南大学医学院生理学系,广东广州510632 [2]广东药学院基础学院,广东广州510006 [3]暨南大学医学院药理学系,广东广州510632

出　　处：《中国病理生理杂志》2015年第12期2113-2119,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81272223)

摘　　要：目的:探讨Cl C-3氯通道是否为IK1钾通道的调节靶点,重点研究鼻咽癌细胞IK1钾通道对Cl C-3氯通道功能及蛋白表达的影响。方法:采用siRNA转染技术抑制低分化鼻咽癌上皮细胞(CNE-2Z)IK1基因的表达;real-time PCR技术检测Cl C-3 mRNA的表达;Western blot检测Cl C-3的蛋白表达;细胞免疫荧光结合激光共聚焦显微镜技术检测Cl C-3和IK1蛋白在细胞内分布;全细胞膜片钳记录细胞氯电流。结果:IK1 siRNA可以成功转染CNE-2Z细胞,有效抑制鼻咽癌细胞IK1钾离子通道的表达;用IK1 siRNA抑制鼻咽癌细胞IK1钾离子通道的表达后,Cl C-3的mRNA表达上调而Cl C-3蛋白却表达减少:在低分化鼻咽癌上皮细胞,低渗刺激可激活氯通道,产生一个较大的氯电流,在成功转染IK1 siRNA的细胞,此氯电流明显减弱。结论:敲低IK1钾离子通道可抑制Cl C-3氯离子通道的表达和功能。AIM： To investigate whether the Cl C-3 chloride channel is an acting target of the IK1 potassium channel,and to study the action of IK1 potassium channel on the functional activities and expression of Cl C-3 chloride channels. METHODS： IK1 gene was silenced by IK1 siRNA in poorly-differentiated nasopharyngeal carcinoma cells（ CNE-2Z）. Real-time PCR and Western blot were used to detect the expression of Cl C-3 at mRNA and protein levels. The distribution of Cl C-3 protein in the cells was observed under confocal immunofluorescence microscope. The chloride current was recorded by the patch-clamp technique. RESULTS： IK1 siRNA was successfully transfected into the CNE-2Z cells and knocked down the expression of IK1 potassium. The mRNA expression of Cl C-3 was increased by the IK1 siRNA. IK1 siRNA inhibited the expression of Cl C-3 protein. A chloride current was activated by hypotonic challenges,and the hypotonicity-induced current was reduced in the cells which successfully transfected with IK1 siRNA. CONCLUSION： The knockdown of IK1 potassium channels inhibits the expression and function of Cl C-3 chloride channel.

关 键 词：CL C-3氯通道 IK1钾通道 鼻咽癌 膜片钳技术 

分 类 号：R730.23[医药卫生—肿瘤] R33-33[医药卫生—临床医学]