BEP2D细胞恶性转化不同时期差异表达基因的筛选

Identification of differentially expressed genes in different malignant transformation phases of BEP2D cell line

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 differentially expressed genes in different malignant transformation phased of BEP2D cell line induced by alpha-particles. Methods Suppression subtractive hybridization(SSH) and cDNA microarray were performed. Results Three suppression subtractive cDNA libraries were constructed.BEP2D cDNA library contained 416 clones,R15H20 cDNA library contained 301 clones,and R15H35 cDNA library contained 586 clones.After confirmed by cDNA microarray,the positive rates in BEP2D cDNA microarray were 90.4%,21.6% and 19.7%;in R15H20 cDNA microarray,8.6%,93.8% and 31.6%;and in R15H35 cDNA microarray,23.5%,18.2%,90.7% when hybridized with probes coming from BEP2D,R15H20 and R15H35 cells,respectively. Conclusions The combination of SSH and cDNA microarray is a rapid and effective method for high throughout screening and identification of differentially expressed genes in different samples.The cDNAs confirmed by cDNA microarray may be the relative genes that associate with malignant transformation of bronchial epithelial cells induced by alpha-particles.