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 DISSENMILATORY MALIGNANT TRANSFORMED HUMAN BRONCHIAL EPITHELIAL CELLS INDUCED BY ALPHA-PARTICLE RADIATION

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 2 0代转化细胞R15Hp2 0的cDNA为tester,BEP2D和R15Hp35细胞的cDNA混合后为driver)有 30 1个克隆 ,C差减文库 (R15Hp35细胞的cDNA为tester,BEP2D细胞的cDNA为driver)有 5 86个克隆。,对文库中 70个cDNA克隆单向测序后发现 :6 1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。

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.