^(125)I-脱氧尿嘧啶核苷对重组质粒pcDNAEgr-IFNγ体外辐射诱导表达的作用  

Experimental study on vitro radiation inducible expression of recombinant plasmid pcDNAEgr-IFNγ by ionizing radiation of ^(125)I-UdR

作　　者：赵敬国[1] 杨巍[2] 孙婷[3] 张巨[1] 李勇[4] 高凤彤[1] 

机构地区：[1]吉林大学中日联谊医院,吉林长春130033 [2]吉林大学卫生部放射生物学重点实验室 [3]吉林大学基础医学院生物化学教研室 [4]哈尔滨医科大学附属第一医院核医学科

出　　处：《中国老年学杂志》2007年第6期501-503,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(30600160)

摘　　要：目的研究125I-脱氧尿嘧啶核苷对重组质粒pcDNAEgr-IFNγ在体外培养B16细胞中的辐射诱导表达作用。方法以脂质体介导法将pcDNAEgr-IFNγ重组质粒转染B16细胞,48h后,于培养液中加入125I-UdR,使其终放射性浓度分别为0(空白对照)、1、5、10、20、50kBq/ml,24h后,采用ELISA法定量检测细胞上清中IFNγ。结果转染组细胞IFNγ蛋白表达随着细胞培养液中125I-UdR浓度的增高而增高,放射性浓度为5、10、20、50kBq/ml组IFNγ的蛋白表达明显高于0kBq/ml组(P<0.05~0.001),其中50kBq/ml组表达量最高,为0kBq/ml组的3.66倍;加入50kBq/ml125I-UdR后6h上清中IFNγ表达量即明显升高(P<0.05~0.001),并随时间延长逐渐增加,照后48h达峰值,表达量为未加入125I-UdR时的6.51倍。结论pcDNAEgr-IFNγ重组质粒在125I-UdR诱导下可有效表达IFNγ,并且其辐射诱导IFNγ基因表达增强特性具有一定的量效和时程规律。Objective To study vitro radiation inducible expression of pcDNAEgr-IFNγrecombinant plasmids by ionizing radiation of ^125I-UdR in B16 cells. Methods pcDNAEgr-IFNγ recombinant plasmid packaged with lipofecfin was stably transfected into B16 cells. 48 h later, the cells were added ^125I-UdR by 0, 1,5, 10, 20 and 50 kBp/nd. 24 h later, the concentration of IFNγ in B16 cells supematant was detected by ELISA. Results IFNγ protein levels with 5, 10, 20, 50 kBq/nd doses of ^125 I-UdR groups were obviously higher than with 0 kBq/nd group （P 〈 0.05- 0.001 ）, IFNγ levels were dose-dependant and the highest level in 50 kBq/nd group was 3.66 times of that of 0 kBq/nd group; IFNγ levels at 6 h in 50 kBp/nd group were obviously higher than that of 0 kBq/nd group （P 〈 0.05 - 0.001 ）, increasing with time, and the highest level at 48 h was 6.51 times of that of 0 kBq/ml group. Conclusions pcDNAEgr-IFNγ induced by ionizing radiation of ^125I-UdR could effectively express IFNT and IFNγ levels are dose-dependant and time-dependant.

关 键 词：B16细胞 基因-放射治疗 脱氧尿苷 碘放射性同位素 

分 类 号：Q786[生物学—分子生物学]