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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 ,探索肿瘤治疗新方法提供了一种实用的细胞模型
In order to further study the mechanism of DNA damage killing tumor cells, to provide a valuable cell model for enhancing the therapeutic effect of tumor, an antisense cDNA was constructed to efficiently express the carboxyl-terminal 10 kD fragment of ATM gene, and was constructed by transfection and screening Cell line U937 ASPI3K stably expressing antisense ATM PI3K cDNA. The results showed that the constructed PI3K cDNA of ATM was sequenced and proved to be reversely located in the expression vector pZeoSV2 (+). After transfected and screened, U937 ASPI3K cell line stably expressing pZeoSV (+) ATM PI3K and U937 pZeoSV (-) cell line, the former ATM protein expression was extremely inhibited, the latter, and the high expression of the U937ATM protein of lymphoma cell line U937ATM Affected. In order to elucidate the role of checkpoint in DNA damage signal transduction pathway and to explore new methods of cancer therapy, this experiment provides a practical cell model