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	一种检测microRNA表达的微阵列芯片的研制及应用
	骆明勇 ; 田志刚 ; 徐智 ; 张亮 ; 王应雄 ; 程京 ; LUO Ming-Yong ; TIAN Zhi-Gang ; XU Zhi ; ZHANG Liang ; WANG Ying-Xiong ; CHENG Jing
	2010-06-09 ; 2010-06-09
关键词	microRNA 微阵列 聚类分析 RT-PCR microRNA, microarray, clustering, RT-PCR Q789
其他题名	Construction and Application of a Microarray for Profiling MicroRNA Expression
中文摘要	MicroRNA(miRNA)是一组内源性的非编码RNA,主要功能是采用降解靶mRNA和抑制靶mRNA的翻译两种作用方式调控基因的表达.研究表明miRNA与动植物的发育、细胞生长分化和凋亡、脂肪代谢以及人类重大疾病密切相关.检测特定状态下的组织或细胞的miRNA表达谱对深入研究miRNA的生物学功能具有重要的意义.研制出了一种检测miRNA表达的微阵列芯片,实验结果显示该芯片具有良好的重复性、灵敏度和特异性.利用这个芯片检测了人脑、肝脏和心脏等组织以及HeLa、HepG2和HL60等细胞株的miRNA表达谱,聚类分析结果显示,miRNA表达模式具有组织特异性,不同个体的同一组织的miRNA表达谱可以聚类在一起.最后,用RT-PCR(reversetranscription-PCR)技术检测了不同组织和细胞株中miR-122a、miR-9和miR-124a的表达,结果显示它们分别在肝脏或脑组织中特异高表达,和芯片结果一致.; MicroRNAs (miRNAs) are a class of endogenous non-coding RNA, which regulate target gene expression via mRNA degradation and translational repression. MicroRNA has been linked to the development of plants and animals, to cell growth and apoptosis, to fat metabolism and other significant physiological processes. Misregulation of microRNA function might contribute to human diseases. Profiling microRNA expression will facilitate the study of biological functions of microRNAs. To detect microRNA expression, a high-density oligonucleotide microarray was constructed, which contains oligonucleotides corresponding to 313 human microRNAs, 261 mouse microRNAs, 196 rat microRNAs and 122 predicted microRNAs. Firstly, high molecular weight RNA was removed from total RNA by polyethylene glycol (PEG) precipitation, and low molecular weight RNA (LMW RNA) was obtained. Then LMW RNA was directly labeled based on a published method that uses T4 RNA ligase to couple LMW RNA to a fluorescence modified dinucleotide. By using a special spotting solution in combination with a swirling hybridization method, the performance of the microRNA microarray was greatly improved. The results of the microRNA microarray experiments indicated its good reproducibility, sensitivity and specificity. It was also showed that the signal of the microRNA microarray was derived from mature microRNAs, but not from their precursors. Using the microarray, microRNA expressions in human brain, heart, liver and in HeLa, HepG2, HL60 cells were profiled. The results revealed a good overall concordance with previously published data. Clustering analysis showed the tissue specificity pattern of microRNA expression, and the same tissue from different individuals was clustered together. Then the expression of miR-122a, miR-9 and miR-124a in human tissues and cell lines with RT-PCR (reverse transcription-PCR) were validated. The results which showed the liver-specific expression of miR-122a and the brain-specific expression of miR-9 and miR-124a were consistent with the microarray results.; 国家科技攻关项目资助(2005BA711A05-04).~~
语种	中文 ; 中文
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/58305]
专题	清华大学
推荐引用方式 GB/T 7714	骆明勇,田志刚,徐智,等. 一种检测microRNA表达的微阵列芯片的研制及应用[J],2010, 2010.
APA	骆明勇.,田志刚.,徐智.,张亮.,王应雄.,...&CHENG Jing.(2010).一种检测microRNA表达的微阵列芯片的研制及应用..
MLA	骆明勇,et al."一种检测microRNA表达的微阵列芯片的研制及应用".(2010).

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