南方医科大学学报2017，Vol.37Issue(2)：256-260,5.DOI:10.3969/j.issn.1673-4254.2017.02.19
Brugada综合征SCN5A基因G1712C突变的功能分析
Functional analysis of a novel SCN5A mutation G1712C identified in Brugada syndrome
摘要
Abstract
Objective To elucidate the molecular and electrophysiological mechanisms of Brugada syndrome through functional analysis of a novel SCN5A gene mutation G1712C.Methods A recombinant plasmid pRc/CMV-hH1 containing the mutant human cardiac sodium channel α subunit (hH1) cDNA was constructed using in vitro PCR-based site-directed mutagenesis technique.LipofectamineTM 3000 was used to transfect the plasmid DNA into HEK293 cell line to induce stable expression of Na+ channel β1-subunit,and the positive colonies were selected by screening with G418.The standard liposome method was used to transiently transfect HEK293 cells with either the wild-type or mutant Na + channel subunits (hH1 and mhH1,respectively),and the macroscopic Na+ currents were recorded using whole-cell patch-clamp technique.Data acquisition and analysis,generation of voltage commands and curve fitting were accomplished with EPC-10,PatchMaster and IGOR Pro 6.0.Results An HEK293 cell line that stably expressed Na + channel β1-subunit was successfully established.After transient transfection with the WT subunit,large Na+ currents were recorded from the stable β1-cell line.Transient transfection with the G1712C subunit,however,did not elicit a Na + current in the cells.Conclusion Compared with normal Na + channel,the wild-type channel exhibits a similar sodium current.The characteristic kinetics of sodium channel of WT-hH1 was identical to that in normal cardiac muscle cell,and the missense mutation (G1712C) in the P-loop region of the domain Ⅳ may have caused the failure of sodium channel expression.关键词
Brugada综合征/SCN5A基因/G1712C/钠电流Key words
Brugada syndrome/SCN5A/G1712C/sodium current引用本文复制引用
陈燕玉,刘深荣,谢亮真,朱庭延,陈益臻,邓晓江,孟素荣,彭健..Brugada综合征SCN5A基因G1712C突变的功能分析[J].南方医科大学学报,2017,37(2):256-260,5.基金项目
广东省科技计划项目(2013B021800140) （2013B021800140）
广州市科技计划项目(201300000146) （201300000146）
