α粒子诱发人支气管上皮细胞恶性转化不同时期差异表达基因cDNA文库的构建

doi:10.3969/j.issn.1004-616x.2002.01.001

癌变·畸变·突变 ›› 2002, Vol. 14 ›› Issue (1): 1-005.doi: 10.3969/j.issn.1004-616x.2002.01.001

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α粒子诱发人支气管上皮细胞恶性转化不同时期差异表达基因cDNA文库的构建

范保星;张开泰;谢　玲;项小琼;吴德昌

军事医学科学院放射医学研究所分子毒理研究室,北京　100850

收稿日期:2001-02-12 修回日期:2001-06-26 出版日期:2002-01-30 发布日期:2002-01-30
通讯作者: 范保星

CONSTRUCTION OF DIFFERENTIALLY EXPRESSED cDNA LIBRARIES FROM DISSENMILATORYMALIGNANT TRANSFORMED HUMAN BRONCHIAL EPITHELIAL CELLS INDUCED BY ALPHAPARTICLE RADIATION

FAN Bao-xing;ZHANG Kai-tai; XIE Ling; XIANG Xiao-qiong; WU De-chang

Department of Molecular Toxicology, Beijing Institute of Radiation Medicine, Beijing 100850, China

Received:2001-02-12 Revised:2001-06-26 Online:2002-01-30 Published:2002-01-30
Contact: FAN Bao-xing

摘要/Abstract

摘要： 目的: 建立α粒子诱发人支气管上皮细胞恶性转化不同时期差异表达基因的文库。方法:抑制消减杂交法(SSH) 。结果:建立了3 个人支气管上皮细胞恶性转化不同时期差异表达基因的cDNA 文库。其中,A 差减文库(永生化人支气管上皮细胞BEP2D 的cDNA 为tester ,α粒子照射BEP2D 细胞后35 代恶性转化细胞R15Hp35 的cDNA 为driver) 有416 个克隆,B 差减文库(α粒子照射BEP2D 细胞后20 代转化细胞R15Hp20 的cDNA 为tester ,BEP2D 和R15Hp35 细胞的cDNA 混合后为driver) 有301 个克隆,C 差减文库(R15Hp35 细胞的cDNA 为tester , BEP2D 细胞的cDNA 为driver) 有586 个克隆。,对文库中70 个cDNA 克隆单向测序后发现:61 个cDNA 为己知基因,9 个cDNA 在GenBank 中无法查到对应的同源序列,可能代表了新基因。结论:3 个差减文库的cDNA 可能代表了α粒子诱发人支气管上皮细胞恶性转化不同时期差异表达的基因,此为进一步研究α粒子诱导肺癌发生的分子机制奠定了基础。

关键词: 抑制消减杂交, BEP2D 细胞, 细胞转化

Abstract: Purpose : To construct differentially expressed cDNA libraries from different malignant transformed human bronchial epithelial cells induced by alpha-particle radiation. Methods : Suppression subtractive hybridization (SSH) . Results : Three differentially expressed cDNA libraries were constructed from different malignant transformed human bronchial epithelial cells. The number of clones is 416 in A subtraction library (The cDNAs of BEP2D cells as tester and R15Hp35 cells as driver) , 301 in B subtraction library ( The cDNAs of R15Hp20 cells as tester and R15Hp35 and BEP2D cells mixed together as driver. ) and 568 in C subtraction library (The cDNAs of R15Hp35 cells as tester and BEP2D cells as driver) . After 70 cDNAs were sequenced and analyzed , 61 cDNAs were found to be known genes , and 9 cDNAs were found to be novel ones. Conclusion : The cDNAs of three subtraction libraries may represent differentially expressed cDNA of different malignant transformed human bronchial epithelial cells induced by alpha-particle radiation.The data provide a basis for further investigation of the molecular mechanism of lung cancer induced by alpha-particle radiation.

Key words: Suppression subtractive hybridization, BEP2D cell, cell transformation

中图分类号:

R811. 5

范保星, 张开泰, 谢　玲, 项小琼, 吴德昌. α粒子诱发人支气管上皮细胞恶性转化不同时期差异表达基因cDNA文库的构建[J]. 癌变·畸变·突变, 2002, 14(1): 1-005.

FAN Bao-xing, ZHANG Kai-tai, XIE Ling, XIANG Xiao-qiong, WU De-chang. CONSTRUCTION OF DIFFERENTIALLY EXPRESSED cDNA LIBRARIES FROM DISSENMILATORYMALIGNANT TRANSFORMED HUMAN BRONCHIAL EPITHELIAL CELLS INDUCED BY ALPHAPARTICLE RADIATION[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2002, 14(1): 1-005.

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α粒子诱发人支气管上皮细胞恶性转化不同时期差异表达基因cDNA文库的构建

CONSTRUCTION OF DIFFERENTIALLY EXPRESSED cDNA LIBRARIES FROM DISSENMILATORYMALIGNANT TRANSFORMED HUMAN BRONCHIAL EPITHELIAL CELLS INDUCED BY ALPHAPARTICLE RADIATION

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