人心房肌Kv1.5钾通道稳定表达细胞系的建立  

Establishment of a cell line stably expressing human cardiac Kv1.5 potassium channel

作　　者：赵志敬[1] 王晓斌[2] 穆润花[3] 刘毅[2] 孙冬冬[1] 李志超[2] 王海昌[1] 董明清[2] 

机构地区：[1]第四军医大学西京医院心血管内科 [2]第四军医大学基础部病理生理学教研室 [3]西京医院妇产科,陕西西安710033

出　　处：《第四军医大学学报》2007年第3期217-219,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30400155)

摘　　要：目的:建立稳定表达人心房肌细胞Kv1.5钾通道的细胞系.方法:将编码人心房Kv1.5钾通道的hKv1.5基因转染HEK293细胞,用1g/LG418筛选2wk后采用膜片钳方法筛选具有抗G418抗性的细胞克隆,电流记录模式为全细胞记录.结果:hKv1.5通道基因被成功转入HEK293细胞并稳定表达;稳定表达的hKv1.5电流是具有明显电压依赖性和超快激活特点的外向电流,其电流大小为(3.20±0.60)nA,其激活电压在-30mV左右.结论:成功构建稳定表达人心房Kv1.5钾通道蛋白的HEK293细胞系,其电流特性与人心房肌IKur相似,可以作为体外研究人心房IKur的细胞模型.AIM： To establish a cell line stably expressing human cardiac Kvl. 5 potassium channel. METHODS： Human cardiac hKvl. 5 gene was transfected into HEK 293 cells. G418 at 1 mg/mL was used to select the transfectod ceUs. After 2 weeks for selection, the anti-G418 cell clones were further identified with patch-clamp method, and whole cell currents were recorded. RESULTS： hKv1. 5 gene was successfully transfected into HEK 293 cells. The expressed hKvl. 5 currents were ultra-rapid outward currents with obvious voltage-dependence. Its average amplitude was （3.20 ± 0.60） nA and its activation voltage was about -30 mV. CONCLUSION： The cell line stably expressing human atrial Kvl. 5 channels has been successfully established, which has the similar characteristics of human atrial IKur. The established cell line can be used as a cell model for studying in vitro human atrial IKur.

关 键 词：心肌/细胞学 钾通道 细胞系 

分 类 号：R331.3[医药卫生—人体生理学]