人红系分化相关基因高表达提高TF-1细胞系存活能力  

Erythroid Differentiation-Associated Gene Overexpression Enhancing the Survival of TF-1 Cell Line

作　　者：郑巍薇[1] 张美江[2] 尹荣华[1] 董小明[3] 詹轶群[1] 杨晓明[1] 李长燕[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所,北京100850 [2]天津大学化工学院,天津300072 [3]北京工业大学生命科学与生物工程学院,北京100022

出　　处：《生物技术通讯》2013年第4期462-466,共5页Letters in Biotechnology

基　　金：国家优秀青年科学基金(81222005);北京市自然科学基金(5122036)

摘　　要：目的:检测人红系分化相关基因(EDAG)对TF-1细胞株存活能力的影响。方法:采用电转染法将过表达EDAG的质粒高效转染TF-1细胞株,通过G418筛选得到过表达EDAG的TF-1细胞稳定株,用MTS法检测过表达EDAG的细胞稳定株在撤除细胞因子后的增殖情况,用流式细胞术检测细胞凋亡。结果:用表达GFP的质粒电转染TF-1细胞株,通过综合评价转染效率和细胞状态,确定最佳转染条件为1350 V电压电击30 ms,电击次数为1次;对得到的细胞稳定株进行检测,其内源EDAG的mRNA和蛋白水平均上调;将细胞稳定株在撤除细胞因子状态下培养,过表达EDAG后细胞存活能力增强、凋亡减少。结论:用电转染方式高效转染TF-1细胞,通过筛选得到过表达EDAG的稳定细胞株,并证实撤除细胞因子后过表达EDAG能提高TF-1细胞系的存活及抗凋亡能力。Objective： To identify the effect of erythroid differentiation-associated gene（EDAG） on the survival of TF-1 cell line. Methods： A TF-1 cell line stably overexpressing EDAG was generated after high-efficiency electrotransfection and G418 screening, and then cell survival was examined by MTS and apoptosis was detected by FACS under the cytokine-free culture. Results： The best condition for transfection was 1350 V, 30 ms, 1 pulse. Both the mRNA and protein level of EDAG were increased in the constructed cell line, EDAG expressing cells showed an increased survival ability and lower apoptosis under the culture without cytokines. Conclusion： An EDAG-overexpressing TF-1 stable cell line was generated and EDAG enhanced the survival of TF-1 cell line under cytokine-free conditions.

关 键 词：人红系分化相关基因 TF-1细胞 存活 凋亡 

分 类 号：Q25[生物学—细胞生物学] R457[医药卫生—治疗学]