稳定表达反义ATM/PI3K细胞系U937-ASPI3K的建立  

Establishment of the Cell Line U937-ASPI3K with Stable Expression of Antisense ATM/PI3K

作　　者：周剑锋[1] 刘文励[1] 汤屹[1] 戴琪琳 孙岚[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院内科,武汉430030

出　　处：《中国实验血液学杂志》2001年第1期22-25,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助 编号 3 980 0 149~~

摘　　要：为进一步研究DNA损伤杀灭肿瘤细胞的机制 ,为增强肿瘤治疗疗效提供一种有价值的细胞模型 ,构建了高效表达ATM基因编码羧基端 10 0kD功能片段的反义cDNA ,通过转染和筛选建立稳定表达反义ATM PI3KcDNA的细胞系U937 ASPI3K。结果显示 ,构建的ATM PI3KcDNA经测序证明反向定位于表达载体 pZeoSV2 (+ )中。经转染和筛选得到稳定表达 pZeoSV(+ ) ATM PI3K的细胞系U937 ASPI3K和对照细胞系U937 pZeoSV (- ) ,前者ATM蛋白表达极度受抑制 ,后者以及淋巴瘤细胞系U937ATM蛋白的高表达未受影响。本实验为阐明细胞周期关卡 (checkpoint)在DNA损伤信号传导通路中的作用机制 。In order to provide a valuable cell model for studying the mechanism of DNA damage to kill tumor cells and enhance the curative effect, an antisense cDNA, which effectively expressed ATM gene encoding carboxyl terminal 100 kD domain fragment, was constructed, and a cell line U937-ASPI3K, which stably expressed the antisense ATM/PI3K cDNA was established through transfection and Zeocin selection. The results showed the constructed ATM/PI3K cDNA is in the opposite orientation in expression vector pZeoSV2(+) through examination of gene sequence. The cell line U937-ASPI3K was total loss of constitutive ATM protein, while the ATM abundance was not influenced in both the control cell lines U937-pZeoSV(-) and U937. The present experiment provided a practical cell model to elusidate mechanism of cell cycle checkpoints in DNA damage signal transduction pathway and search a new approach for tumor therapy.

关 键 词：反义ATM/PI3K U937-ASPI3K细胞系 U937细胞系 基因突变 共济失调性毛细血管扩张症 

分 类 号：R543.7[医药卫生—心血管疾病] R730.5[医药卫生—内科学]