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| Pkm2-shRNA真核表达载体的构建和对急性早幼粒细胞白血病耐药细胞株的作用研究 | |
| 引用本文： | 孙明莉,王冠军,李江,崔久嵬,张爱丽,王中南,李晓萌,李薇. Pkm2-shRNA真核表达载体的构建和对急性早幼粒细胞白血病耐药细胞株的作用研究[J]. 中国实验血液学杂志, 2010, 18(1): 85-89 |
| 作者姓名： | 孙明莉 王冠军 李江 崔久嵬 张爱丽 王中南 李晓萌 李薇 |
| 作者单位： | 1. 吉林大学第一医院肿瘤中心,吉林长春,130021 2. 吉林大学口腔医院口腔修复科,吉林长春,130041 3. 东北师范大学生命科学学院,吉林长春,130024 |
| 基金项目： | 吉林省科技厅社会发展重大项目,编号20050412-1 |
| 摘 要： | 本研究旨在构建M2型丙酮酸激酶(M2-pyruvate kinase;PKM2)的小发夹结构RNA(shRNA)的真核表达载体,研究特异性沉默pkm2基因对急性早幼粒细胞白血病(APL)耐药性的影响。设计pkm2基因的3个特异shRNA序列,利用基因重组技术将其克隆到含有U6启动子的PBSU6载体上,通过酶切和测序等方法鉴定重组质粒的正确性。利用Western blot方法检测了pkm2-shRNA对急性早幼粒细胞白血病耐药细胞株NB4R2细胞内源性M2-PK蛋白表达的影响,通过NBT还原实验检测了pkm2基因沉默后NB4R2细胞分化情况。结果表明:成功构建了3个有效特异的pkm2-shRNA,在蛋白水平上证明了其对NB4R2细胞M2-PK基因沉默的有效性。发现干涉M2-PK基因后,可显著地促进耐药细胞NB4R2细胞的分化。结论:DNA载体途径的pkm2-shRNA能促进早幼粒细胞白血病细胞的分化,具有基因靶向药物开发前景。 |
| 关 键 词： | M2型丙酮酸激酶 小发夹结构RNA NB4R2细胞 急性早幼粒细胞白血病 |
Construction of shRNA Eukaryotic Expression Vectors of pkm2 Gene and Their Effect on Drug Resistant Cell Line of Acute Promyelocytic Leukemia |
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| SUN Ming-Li,WANG Guan-Jun,LI Jiang,CUI Jiu-Wei,ZHANG Ai-Li,WANG Zhong-Nan,LI Xiao-Meng,LI Wei. Construction of shRNA Eukaryotic Expression Vectors of pkm2 Gene and Their Effect on Drug Resistant Cell Line of Acute Promyelocytic Leukemia[J]. Journal of experimental hematology, 2010, 18(1): 85-89 | |
| Authors: | SUN Ming-Li WANG Guan-Jun LI Jiang CUI Jiu-Wei ZHANG Ai-Li WANG Zhong-Nan LI Xiao-Meng LI Wei |
| Affiliation: | ( Cancer Center, The First Hospital, Jilin University, Changchun 130021, Jilin Province, China ; 1 Department of Oral and Maxillofacial Surgery, Stomatologic Hospital, Jilin University, Changchun 130041, Jilin Province, China ; School of Life Sciences, Northeast Nor- real University, Changchun 130024, Jilin Province, China) |
| Abstract: | This study was aimed to construct the shRNA eukaryotic expression vectors of M2-pyruvate kinase gene(pkm2) and to investigate the effects of pkm2 gene interference on the drug resistance of acute promyelocytic leukemia(APL)cells in vitro.Three specific shRNAs of pkm2 gene were designed and cloned into PBSU6 vector containing a U6 promotor.The constructed plasmids were identified and proved by the restriction sequence analysis.Then the effect of pkm2-shRNA on the protein expression of endogenous PKM2 was det... |
| Keywords: | M2-pyruvate kinase shRNA NB4R2 cell acute promyelocytic leukemia |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |