MicroRNA在六种不同器官肿瘤细胞系中的差异表达  

Differential Expression of MicroRNA in 6 Kinds of Human Cancer Lines from Various Organs

作　　者：马卓娅[1] 汤华[1] 李欣[1] 刘民[1] 吴海东[1] 王晶[1] 

机构地区：[1]天津市生命科学中心实验室,天津医科大学,天津300070

出　　处：《中国肿瘤》2007年第9期714-717,共4页China Cancer

摘　　要：[目的]利用生物芯片技术分析microRNA(miRNA)在六种不同器官肿瘤细胞系中mi-croRNA(miRNA)的表达差异。[方法]将210个(206个miRNA,4个阳性对照)与已知人类和小鼠miRNA互补的序列作为探针,点于玻片上制备寡核苷酸芯片。提取肿瘤细胞(HeLa、MCF-7、A549、HT-29、ES-2、K562)的miRNA,用荧光染料Cy3标记,并与制备好的芯片杂交。用Sca-nArrayTMExpress1.0扫描仪扫描荧光信号,采用ScanArray3.0和Cluster3.0软件分析处理扫描结果。并用Northernblot和RT-PCR方法对芯片检测结果进行验证。[结果]芯片检测到在六种不同器官肿瘤细胞系中,发现115种miRNA存在差异,91种miRNA没有明显差异。其中,miR-21在六种肿瘤细胞表达水平均较高,miR-125b表达水平均较低,let-7在六种细胞系中表达水平较低。miR-17-5p和miR-20a呈集簇表达,在卵巢肿瘤ES-2细胞系中的表达水平高于其它细胞,乳腺肿瘤细胞系MCF-7和宫颈癌细胞系HeLa的miRNA的表达谱聚为一类。[结论]应用生物芯片技术检测细胞中miRNA的表达差异谱是可行的,miRNA可能参与肿瘤的发生发展。[Purpose ] To analyze the differential expression of microRNA（miRNA） in 6 kinds of human cancer cell lines from various organs. [Methods ] The microarray was prepared by spotting 210 oligonucleotides which contains 206 probes complementary with human and mouse miRNA sequence and 4 positive control oligos. MiRNAs from HeLa,MCF-7,A549,HT-29,ES-2,K562 were extracted and labeled with Cy3 and then hybridized to the miRNA microarray. Then, slides were scanned by ScanArray^TM Expressl.0 and images were analyzed by ScanArray3.0 and Cluster3.0. Results were confirmed by Northern blot and RT-PCR. [Results] Different expression profiles were detected in 6 cancer cell lines and 115 miRNAs were differentially expressed,and 91 kinds of miRNAs had no remarkable difference,among which miR-21 expressed with high level,while miR-125b, let-7 expressed with low level. Expression of miR-17-5p and miR-20a revealed cluster, and it was higher in ES-2 than that in other cells. HeLa and MCF-7 were located on a single branch of the dendrogram i,is cluster analys. [Conclusion] It is feasible to detect the miRNA expression profiles with microarrays, miRNA might be involved in carcinogenesis.

关 键 词：寡核苷酸芯片 MICRORNA 肿瘤 表达谱 

分 类 号：R73-354[医药卫生—肿瘤]