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

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.

CONSTRUCTION OF DIFFERENTIALLY EXPRESSED cDNA LIBRARIES FROM DISSENMILATORY MALIGNANT TRANSFORMED HUMAN BRONCHIAL EPITHELIAL CELLS INDUCED BY ALPHA-PARTICLE RADIATION

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.