针对EDAG-1基因的siRNA抑制白血病细胞的生长及IL-8分泌  

Inhibitory effect of siRNA specific for EDAG-1 on growth of leukemia cells and the secretion of IL-8

作　　者：周颖[1] 程勇[2] 肖敏[1] 高宗侠[1] 程志祥[1] 冯定庆[1] 凌斌[1] 

机构地区：[1]安徽省立医院安徽省分子医学重点实验室分子医学研究中心,合肥230001 [2]安徽省立医院放疗科,合肥230001

出　　处：《肿瘤》2008年第7期577-580,共4页Tumor

基　　金：安徽省教育厅自然科学研究资助项目(编号:2006KJ080C)

摘　　要：目的:探讨抑制胚胎发育相关基因-1(embryonic develop-associated gene1,EDAG-1)表达对白血病细胞株生长的影响及其机制。方法:将靶向EDAG-1基因的特异序列连接到逆转录病毒载体中,将脂质体介导的重组质粒转染包装产病毒细胞株PT67;收集细胞上清液转染HEL细胞株,经嘌呤霉素筛选获得稳定抑制EDAG-1基因表达的细胞株;RT-PCR法检测EDAG-1基因的抑制情况,MTT法检测转染细胞的增殖能力,同时采用RT-PCR和ELISA法检测转染后细胞中IL-8的表达水平。结果:成功构建获得靶向EDAG-1基因的反转录病毒载体,筛选获得抑制EDAG-1基因表达的HEL细胞株。EDAG-1基因被抑制的细胞株增殖速度降低,IL-8的分泌水平同时相应减低。结论:沉默EDAG-1基因的表达可以抑制白血病细胞株的增殖和IL-8的分泌,提示EDAG-1基因可能是白血病治疗的一个有效靶点。Objective： To explore the effects of silencing the expression of embryonic develop-associated gene-1 （EDAG-1） by siRNA on the growth of leukemia cell lines and the action mechanisms. Methods：The specific oligonucleotides targeting EDAG-1 were cloned into retroviral vector. The recombinant retroviral vectors were transfected into RetroPack PT67 cell line. The supernatant was collected from the cultured transfected PT67 cells and transfected into HEL cells. The stable transfected HEL cell lines were selected with puromycin. The down-regulation of EDAG-1 expression was determined by RT-PCR. The proliferation of HEL cells was evaluated by MTT assay. The release of IL-8 was analyzed by RT-PCR and ELISA. Results： The recombinant retroviral vectors were successfully constructed. The stable HEL cell line with a persistent knockdown of EDAG-1 gene was obtained by screening. Down-regulation of EDAG-1 expression by siRNA inhibited the proliferation speed of cells and reduced the secretion of IL-8. Conclusion： Silencing the expression of EDAG-1 slowed clown the proliferation of leukemia cell lines and decreased the secretion of IL-8, suggesting that EDAG-1 has the potential to become an effective target for leukemia therapy.

关 键 词：白血病 RNA 小分子干扰 基因 肿瘤抑制 基因 胚胎发育相关 

分 类 号：R73-36[医药卫生—肿瘤]