甲状腺癌细胞系方波电脉冲基因转染条件的探讨  

Effective gene transfer parameters to thyroid cancer using square - wave Electroporation gene delivery technique

作　　者：孙辉[1] 王金国[1] 单纯玉[1] 付言涛[1] 

机构地区：[1]吉林大学中日联谊医院普外科,长春130031

出　　处：《中国地方病防治》2003年第5期268-270,共3页Chinese Journal of Control of Endemic Diseases

摘　　要：目的 探讨方波脉冲基因电转染对人甲状腺癌细胞系的转染条件和效果。方法 选用pEGFP-C1作为外源基因与方波电脉冲相结合,以人甲状腺癌细胞系TA-K为导入对象,探讨方波脉冲基因电转染对人甲状腺癌细胞系转染的脉冲幅度,脉冲时值,脉冲次数和反应体系大小。结果 当脉冲幅度在60V/mm时,开始出现阳性细胞,脉冲时值20ms时,基因转染效率可高达35％,细胞生存分数55％,脉冲时值20ms、次数1次转染效率高、细胞死亡率低。200μl混合液电转染后观察,对细胞生存影响不大。结论 脉冲幅度在60V/mm,脉冲时值20ms,脉冲次数1次,反应体系200μl,可以为人甲状腺癌细胞系的转染提供良好效果。Objective In this studywe measured transfection efficiency in thyroid cancer cell line using square - wave Electroporation gene delivery technique. Methods A plasmid encoding the green fluorescent protein( pEGFP - Cl )was selected to be transfect-ed to thyroid cancer cell line(TA - K) . Emphasis was placed on different electroporation parameters, such as amplitude and duration. Transfertion efficiencies were evaluated under different electroporation conditions. Results 50 - 400μ1 of plasmid - cell mixtures were transferred to a 2 - mm gap cuvette with parallel - plate. 1 - 4 square - wave pulses were applied at amplitudes 0 - 60V/mm, with durations 0 - 40 ms time and 1 Hz frequency. Conclusions 200μl of plasmid - cell mixture, 1 square - wave pulse, amplitude 60V/mm, duration 20 ms time, were effective gene transfer parameters to TA - K.

关 键 词：甲状腺癌 方波电脉冲基因转染 癌细胞 外源基因 

分 类 号：R736.1[医药卫生—肿瘤]