| RNAi重组体抑制白血病K562细胞系中NPM1基因的表达 |
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| 引用本文： | 何鹏,骆展鹏,张伶,杨松,钟晓明,高玉洁. RNAi重组体抑制白血病K562细胞系中NPM1基因的表达[J]. 第三军医大学学报, 2008, 30(10): 938-941 |
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| 作者姓名： | 何鹏 骆展鹏 张伶 杨松 钟晓明 高玉洁 |
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| 作者单位： | 重庆医科大学医学检验系临床血液学教研室,临床检验诊断学省部共建教育部重点实验室,重庆市重点实验室,重庆,400016 |
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| 摘 要： | 目的 构建核仁磷酸蛋白1(nucleophosmin1,NPM1)特异性的RNAi真核表达载体,观察对白血病K562细胞系NPM1表达的抑制作用.方法 采用RT-PCR检测白血病细胞系(THP1和K562)和急性髓系白血病(AML)细胞NPM1 mRNA水平.设计合成2条针对NPM1基因并具有小发夹结构的DNA序列,经退火形成互补双链.再克隆至载体pGenSil-1中构建含NPM1的RNAi重组体,经酶切及测序鉴定后通过脂质体转染K562细胞,同时设立pGenSil-1转染组和pGenSil-1/neg转染组.采用RT-PCR检测各组转染细胞中NPM1的mRNA水平.结果 白血病细胞系和AML细胞均高表达NPM1 mRNA.针对人NPM1基因的RNAi重组体构建成功,并能够稳定转染K562细胞,导致白血病细胞NPM1 mRNA相对表达量下降64%,而pGenSil-1转染组和pGenSil-1/neg转染组未能下调NPM1表达.结论 白血病细胞高表达NPM1基因,其mRNA表达水平能够被靶向NPM1的RNAi重组体特异性地下调.
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| 关 键 词： | 核仁磷酸蛋白1 RNA干扰 白血病 K562细胞 |
| 文章编号： | 1000-5404(2008)10-0938-04 |
| 修稿时间： | 2007-10-19 |
RNA interference silencing nucleophosmin 1 inhibits NPM1 gene expression in K562 cells |
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| HE Peng,LUO Zhan-peng,ZHANG Ling,YANG Song,ZHONG Xiao-ming,GAO Yu-jie. RNA interference silencing nucleophosmin 1 inhibits NPM1 gene expression in K562 cells[J]. Acta Academiae Medicinae Militaris Tertiae, 2008, 30(10): 938-941 |
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| Authors: | HE Peng LUO Zhan-peng ZHANG Ling YANG Song ZHONG Xiao-ming GAO Yu-jie |
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| Affiliation: | HE Peng,LUO Zhan-peng,ZHANG Ling,YANG Song,ZHONG Xiao-ming,GAO Yu-jie(Department of Clinical Hematology,Faculty of Laboratory Medicine,Key Laboratory of Clinical Medical Diagnostics of Education Ministry,Chongqing Medical University,Chongqing 400016,China) |
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| Abstract: | Objective To construct eukaryotic expression vector of RNAi specific to nucleophosmin1(NPM1)and investigate the effect of recombinant plasmid on NPM1 expression in K562 cells.Methods The expression level of NPM1 mRNA was detected by RT-PCR in leukemia cell lines(THP1 and K562),bone marrow or blood samples from 5 patients with acute myelogenous leukemia(AML)and from 5 healthy subjects.A short hairpin RNA targeting NPM1 was synthesized and inserted into the plasmid pGenSil-1.After the recombinant plasmid was ... |
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| Keywords: | nucleophosmin 1 RNA interference leukemia K562 cells |
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