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目的 筛选并鉴定α粒子辐射诱发永生化人支气管上皮细胞 (BEP2D)恶性转化不同时期差异表达的基因。方法 抑制消减杂交 (SSH) ,cDNA芯片。结果 利用SSH构建了BEP2D细胞 3个恶性转化不同时期差异表达基因的文库 ,BEP2D细胞的文库有 4 16个克隆 ,R15H2 0细胞的文库有 30 1个克隆 ,R15H35细胞的文库有 5 86个克隆 ,经cDNAMicroarray验证发现 :BEP2D细胞文库来源的芯片与 3代细胞探针杂交后的阳性信号率为 90 4 %、2 1 6 %和 19 7% ;R15H2 0细胞文库来源的芯片的阳性信号率为 8 6 %、93 8%和 31 6 % :R15H35细胞文库来源的芯片的阳性信号率为 2 3 5 %、18 2 %和 90 7%。结论 联合应用SSH和cDNAMicroarray是筛选和鉴定不同样本中差异表达基因的快速和有效方法 ;经cDNAMicroarray鉴定出的 3个文库中差异表达的cDNA可能代表了α粒子诱发人支气管上皮细胞恶性转化相关的基因。
Objective To screen and identify the genes differentially expressed in malignant transformation of immortalized human bronchial epithelial cells (BEP2D) induced by α particle irradiation. Methods Suppression subtractive hybridization (SSH), cDNA microarray. Results A total of 4 16 clones of BEP2D cells, 30 1 clones of R15H20 cells and 5 86 clones of R15H35 cells were constructed. The results of cDNA microarray showed that the positive signal rates of BEP2D cell library were 90 4%, 21 6% and 19 7% after hybridization with the 3rd generation cell probe. The positive signal rate of the chip derived from R15H20 cell library was 8 6%, 93 8%, and 31 6%. The positive signal rates of the R15H35 cell-derived chip were 23.5%, 18.2% and 90.7%. Conclusions The combined application of SSH and cDNA microarray is a rapid and effective method for screening and identifying differentially expressed genes in different samples. The cDNAs differentially expressed in the three libraries identified by cDNA Microarray may represent the genes involved in α-particle-induced malignant transformation of human bronchial epithelial cells .