山东医药Issue(15)：20-23,4.DOI:10.3969/j.issn.1002-266X.2014.15.007
pGPH1-GFP-RKIP 的构建及其对白血病细胞株 CCRF-CEM 化疗敏感性的影响
Establishment of pGPH1-GFP-RKIP plasmid vector and its effect on sensitivity to chemotherapy of leukemia cell line CCRF-CEM
李青 1多力坤2
作者信息
- 1. 武警兵团指挥部医院，乌鲁木齐830063
- 2. 新疆医科大学第一附属医院
- 折叠
摘要
Abstract
Objective To establish plasmid vectors pGPH-green fluorescent protein ( GFP)-Raf kinase inhibitor pro-tein ( RKIP) expressing RKIP small interfering RNA ( siRNA) and to observe the effect of RKIP RNA interference on sen-sitivity to chemotherapy of pediatric leukemia cells CCRF-CEM.Methods Plasmid RKIP interference vectors pGPH-GFP-RKIP were established and transfected into CCRF-CEM cells by using electroporation technique .RKIP, phosphorylated ex-tracellular regulated protein kinase (p-ERK) and phosphorylated nuclear factor κB inhibitor (p-IκB) levels in the trans-fected cells were detected by using Western blotting .9-nitrocamptothecin (9NC) at various concentrations were used to in-duce CCRF-CEM cells apoptosis , and Western blotting was used to detect the RKIP expression , caspase-3 activities ( apop-tosis rates) were detected by using flow cytometry .Results The pGPH1-GFP-RKIP was successfully established .Trans-fection rates of pGPH1-GFP-RKIP and pGPH1-GFP plasmids were 90.04% and 91.32%, respectively.The expression levels of RKIP were effectively down-regulated and both p-ERR and p-IκB levels were up-regulated in CCRF-CEM cells transfected with pGPH-GFP-RKIP plasmids as compared with the cells transfected with pGPH 1-GFP plasmids ( all P<0.05).RKIP levels in the CCRF-CEM cells exposed to 9NC were up-regulated and the CCRF-CEM cells transfected with pGPH-GFP-RKIP had down-regulated apoptosis rates (all P<0.05).Conclusions RKIP siRNA plasmid vectors are suc-cessfully established .RKIP expression changes may be associated with the sensitivity to chemotherapy of leukemia cells .关键词
Raf激酶抑制蛋白/RNA干扰/儿童白血病/化疗敏感性Key words
Raf kinase inhibitor protein/RNA interference/pediatric leukemia/sensitivity to chemotherapy分类
医药卫生引用本文复制引用
李青,多力坤..pGPH1-GFP-RKIP 的构建及其对白血病细胞株 CCRF-CEM 化疗敏感性的影响[J].山东医药,2014,(15):20-23,4.
