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利用抑制性消减杂交技术鉴定镍暴露大鼠肺组织中的差异表达基因。

Identification of differentially expressed genes in lung tissues of nickel-exposed rats using suppression subtractive hybridization.

机构信息

School of Life Science and Technology, Tongji University, Shanghai, 200092, China.

出版信息

Biol Trace Elem Res. 2011 Nov;143(2):1007-17. doi: 10.1007/s12011-010-8898-4. Epub 2010 Nov 18.

Abstract

Occupational exposure to nickel compound, such as nickel refining, electroplating, and in conjunction with other metals, is harmful to the health, causing respiratory distress, and lung and nasal cancer. In this work, the different gene expression patterns of lung tissues from nickel-exposed rats and controls were investigated. The suppression subtractive hybridization (SSH) method was used to generate two subtracted cDNA libraries with gene transcripts differentially expressed after nickel inducing. Dot-blot hybridizations were used to confirm differential ratios of expression of obtained SSH clones. Out of 768 unique SSH clones, which were chosen randomly from the two subtraction libraries (384 of each), 319 could be verified as differentially expressed. According to blast screening and functional annotation, 28% genes in nickel-induced cDNA library were related to cell differentiation, whereas 21% in driver library were related to oxygen transport. Two novel expressed sequence tags (ESTs; NCBI Accession No. FC809414 and No. FC809411) in nickel-induced cDNA library were obtained. The genes detected in the present study are probably important genes associated with nickel-induced lung cancer.

摘要

职业性接触镍化合物,如镍精炼、电镀,以及与其他金属结合,对健康有害,导致呼吸困难、肺癌和鼻癌。在这项工作中,研究了镍暴露大鼠和对照组肺组织的不同基因表达模式。抑制性消减杂交(SSH)方法用于产生两个差异表达基因转录本的消减 cDNA 文库,这些基因转录本在镍诱导后表达。点杂交杂交用于确认获得的 SSH 克隆的表达差异比率。从两个消减文库(各 384 个)中随机选择的 768 个独特 SSH 克隆中,有 319 个可被验证为差异表达。根据 Blast 筛选和功能注释,镍诱导 cDNA 文库中 28%的基因与细胞分化有关,而驱动子文库中 21%的基因与氧气运输有关。镍诱导 cDNA 文库中获得了两个新的表达序列标签(EST;NCBI 注册号 FC809414 和 FC809411)。本研究中检测到的基因可能是与镍诱导肺癌相关的重要基因。

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