NDRG2基因六种可变剪接体的分型鉴定  

Identification of Altematively Spliced Variants of Human NDRG2 Gene and its Application

作　　者：尹安琪 孙佳辰 张娜娜[2] 药立波[2] 李燕[1,2] 熊利泽[1] 

机构地区：[1]第四军医大学西京医院麻醉科,陕西西安710032 [2]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710032

出　　处：《现代生物医学进展》2016年第20期3835-3838,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81471110)

摘　　要：目的:抑癌基因N-myc下游调节基因2(N-myc down stream regulated gene 2,NDRG 2)在抑制肿瘤的发生及进展过程中具有重要作用,本研究旨在区分并鉴定NDRG2基因mRNA的6种可变剪接体。方法:针对NDRG2基因mRNA6种剪接体外显子的差别设计6对特异性引物。分别从正常人脑组织和胶质瘤细胞U251中提取总RNA,反转录为cDNA,利用高保真酶扩增目的剪接体DNA,并根据PCR扩增产物电泳条带的有无和大小初步判断剪接体的型别,最后将PCR产物胶回收进行测序鉴定。结果:利用本实验中设计的引物可以将人脑组织和U251胶质瘤细胞中的NDRG 2基因mRNA剪接体经行分型鉴定,确定了分型引物的可行性。人脑组织和U251细胞中均含有A、B、D和E四种剪接体。结论:利用本研究设计的特异性引物进行PCR扩增,可以判断某种细胞或组织中表达的NDRG2mRNA分子剪接体的型别。有助于未来深入研究抑癌基因NDRG2在肿瘤发生发展中的抑制作用与不同剪接体的表达相关性。Objective： Tumor suppressor gene N-myc 2 （N-myc down stream regulated gene 2, NDRG2）plays important role in suppressing the tumorigenesis and progressing. Our research aims to differentiate and identify six variants ofNDRG2 mRNA. Methods： Based on the differences of six variants, we designed six specific primers. We extracted total RNA from normal brain tissue and glioma cell line U251 respectively, using RT-PCR to get the targeted eDNA. Then we differentiate the six variants by electrophoresis. Finally we recollected the RT-PCR products to test the sequencing. Results： With the specific primers we confirmed that both normal human brain tissue and glioma cell line U251 contain the A, B, D, and E, four kinds of variants. Conclusion： With the specific primers we designed, we can tell the differences of NDRG2 variants expression in different cells or tissues, which help to further explore the relationship between the NDRG2 variants expression and the tumor suppressing effect.

关 键 词：N-myc下游调节基因2 剪接体 基因表达 PCR 

分 类 号：Q78-6[生物学—分子生物学] R34[医药卫生—基础医学]