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氡染毒小鼠外周血细胞差异表达基因的筛选与鉴定
  • ISSN号:0254-5098
  • 期刊名称:《中华放射医学与防护杂志》
  • 时间:0
  • 分类:R735.7[医药卫生—肿瘤;医药卫生—临床医学] R135.1[医药卫生—劳动卫生;医药卫生—公共卫生与预防医学]
  • 作者机构:[1]苏州大学附属第二医院呼吸科,215004, [2]苏州大学放射医学与公共卫生学院卫生毒理学教研室
  • 相关基金:国家自然科学基金资助项目(30371226,30671784);教育部博士学科点专项科研基金资助项目(20050285009)
中文摘要:

目的在建立氡染毒小鼠模型的基础上,筛选和鉴定氡染毒小鼠外周血细胞差异表达基因。方法采用HD-3型多功能氡室对BALB/c小鼠动态吸入染毒,建立实验动物模型。按照小鼠吸入氡及其子体的累积剂量分为实验组(累积剂量105WLM)和对照组(室内本底浓度,1WLM)。采用RNA转录物5’末端启动机制技术(SMART)和抑制性消减杂交技术(SSH)构建正、反向消减eDNA文库,选取含有插入eDNA片段的克隆进行反向Northern杂交筛选上调或下调的阳性克隆。克隆的测序结果进行同源性检索和生物信息学分析。结果在成功构建了消减cDNA文库基础上,挑取白斑390个,获得312个含有插入片段的单克隆,进行反向Northern杂交,分析基因的上调或下调水平。选取差异表达克隆41个进行测序及生物信息学分析,最终获得10条已知功能或有注释的差异基因片段和3条未知的表达序列标签(EST)。筛选出的差异表达基因涉及氧化损伤、细胞增殖和肿瘤发生等多方面的功能。结论氡染毒可引起一系列基因表达的上调或下调,利用SSH技术可以筛选到氡染毒的差异表达基因,为探讨氡损伤机制提供参考。

英文摘要:

Objective To screen and identify the differential expression genes on peripheral blood cells of mice based on the experimental animal model of radon exposure. Methods BALB/e mice were exposed in a type HD-3 multifunctional radon-room, with the accumulative doses of radon-exposure group at 105 WLM and control group at 1 WLM. Total RNA was extracted from peripheral blood cells and the methods of SMART for dscDNA synthesis and SSH for gene screening was applied. With the construction of the eDNA library enriched with differentially expressed genes, the pMD 18-T plasmid containing LacZ operator at the multiple cloning site was used to allow a blue-white screening. The TA clones were amplified by nested PCR and the reverse Northern blot was used to identify up and down regulation of the clones. The differently expressed eDNA was then sequenced and analyzed. Results The subtracted eDNA libraries were successfully constructed. A total of 390 recombinant white colonies were randomly selected. Among the 312 eDNA monoclones selected from both forward- and reverse-subtracted libraries,41 clones were chosen to sequence for their differential expressions based on reverse Northern blot. Among the 41 sequenced clones, 10 clones with known function/annotation and 3 new ESTs with the GenBank accession numbers were obtained. Most of the known function/annotation genes were revealed to be related with cell proliferation, metabolism, cellular apoptosis and carcinogenesis. Conclusions The animal model of radon exposure was established and the cDNA library of peripheral blood cells was successfully constructed. Radon exposure could up- and clown-regulate a series of genes. Differentially expressed genes could be identified by using SSH technique and the results may help exploring mechanisms of random exposure.

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期刊信息
  • 《中华放射医学与防护杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中华医学会
  • 主编:
  • 地址:北京市德外新康街2号
  • 邮编:100088
  • 邮箱:cjrmp@cjrmp.sina.net
  • 电话:010-62389620
  • 国际标准刊号:ISSN:0254-5098
  • 国内统一刊号:ISSN:11-2271/R
  • 邮发代号:18-93
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:11741