高精度原子力显微镜显示磁性反义探针与SK-Br-3肿瘤细胞mRNA核苷酸连接  

Connection of Magnetic Antisense Probe with SK-Br-3 Oncocyte mRNA Nucleotide Detected by High Resolution Atomic Force Microscope

作　　者：谭书德[1,2] 欧阳羽 李信友[1] 文明[1] 李少林[1] 

机构地区：[1]重庆医科大学附属第一医院放射科,重庆400016 [2]重庆市綦江县人民医院CT室

出　　处：《生物医学工程学杂志》2011年第3期442-445,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30070230,30940021);重庆市自然科学基金资助项目(2008BB5209);重庆市卫生局医学科研基金资助项目(062025);重庆医科大学优秀博士基金资助项目(2006-182),重庆医科大学附属第一医院科研基金资助项目(YXJJ2009-08)

摘　　要：为了将高精度原子力显微镜(AFM)用于显示超顺磁性氧化铁标记的c-erbB2癌基因反义寡脱氧核苷酸探针(磁性反义探针)与SK-Br-3肿瘤细胞mRNA核苷酸的连接,我们在磁性反义探针转染SK-Br-3肿瘤细胞基础上,用AFM对转染后的肿瘤细胞进行观察,并同时对转染后的肿瘤细胞进行蛋白表达检测及MRI成像,以进一步证实AFM的观察结果。从AFM显示的磁性反义探针转染SK-Br-3肿瘤细胞后单个细胞的全貌图及局部放大图发现,探针中反义寡脱氧核苷酸中的脱氧胞嘧啶核苷酸闭环与肿瘤细胞mRNA嘌呤核苷酸环相连接;此外,磁性反义探针能特异性抑制SK-Br3细胞c-erbB2的蛋白表达,MRI显示磁性反义探针转染SK-Br-3肿瘤细胞的信号强度最低(P<0.05)。实验表明,AFM可以清楚显示磁性反义探针与SK-Br-3肿瘤细胞核mRNA核苷酸的连接。The present paper is aimed to detect superparamagnetic iron oxide labeled c-erbB2 oncogene antisense oligonucleotide probe（magnetic antisense probe） connected with SK-Br-3 oncocyte mRNA nucleotide by high resolution atomic force microscope（AFM）.We transfected SK-Br-3 oncocyte with magnetic antisense probe,then observed the cells by AFM with high resolution and detected protein expression and magnetic resonance imagine（MRI）.The high resolution AFM clearly showed the connection of the oligonucleotide remote end of magnetic antisense probe with the mRNA nucleotide of oncocyte.The expression of c-erbB2 protein in SK-Br3 cells were highly inhibited by using magnetic antisense probe.We then obtained the lowest signal to noise ratio（SNR） of SK-Br-3 oncocyte transfected with magnetic antisense probe by MRI（P0.05）.These experiments demonstrated that the high resolution AFM could be used to show the binding of magnetic antisense probe and SK-Br-3 mRNA of tumor cell nuclear.

关 键 词：原子力显微镜 反义寡脱氧核苷酸 超顺磁性氧化铁 SK-Br-3肿瘤细胞株 

分 类 号：R730.2[医药卫生—肿瘤]