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目的应用基因芯片分析X射线照射对来源于人正常淋巴母细胞系的AHH-1基因表达的影响。方法照射组采用基因芯片分析0.1、0.5、2.0、5.0 Gy X射线(设为相应的剂量组)照射AHH-1后培养20 h基因表达谱的改变,另设1组未照射组为对照组。将照射组与对照组细胞的基因表达量差异比值>2.0或<0.5确定为有效差异表达基因。用逆转录聚合酶链反应(RT-PCR)和实时聚合酶链反应(Real-time PCR)方法验证部分差异表达基因。结果照射组共同表达上调的基因有19个,共同表达下调的基因有22个。不同剂量组基因差异表达的倍数变化幅度较大,从下调0.05倍到上调13.12倍,但大部分在下调0.30~0.50倍和上调2.00~3.00倍,其中共济失调–毛细血管扩张突变基因在0.1和0.5 Gy剂量组表达上调,细胞周期依赖性激酶抑制1A基因(CDKN1A)在2.0和5.0 Gy剂量组表达上调。RT-PCR检测结果与基因芯片结果相一致,包括CDKN2A基因、肿瘤坏死因子受体超级家族19基因(TNFRSF19)、核糖核酸酶7基因(RNASE7)和早期生长反应因子3基因(EGFR3),Real-time PCR证实照射后TNFRSF19的表达下降。结论不同剂量X射线照射对AHH-1细胞多种功能基因的表达产生了影响,照射后差异表达基因参与不同的生物学过程。
Objective To analyze the effect of X-ray irradiation on the expression of AHH-1 gene in human normal lymphoblastoid cell line using cDNA microarray. Methods The gene expression profile of AHH-1 cultured in 0.1, 0.5, 2.0, and 5.0 Gy X-rays (set as the corresponding dose group) was analyzed by gene chip after 20 h of gene expression change, and another group of non-irradiated group was set as the control group. The ratio of gene expression of irradiation group and control group was> 2.0 or <0.5, which was determined as a valid differentially expressed gene. Some differentially expressed genes were verified by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR. Results There were 19 genes up-regulated in co-expression group and 22 genes co-expressed down-regulated in irradiation group. The fold-change of gene differential expression in different dose groups was larger, ranging from 0.05-fold up to 13.12-fold, but most of them were down-regulated from 0.30 to 0.50-fold and 2.00- to 3.00-fold higher, with ataxia-telangiectasia mutated in 0.1 And 0.5 Gy dose group, up-regulated expression of cyclin-dependent kinase 1 (CDKN1A) gene in 2.0 and 5.0 Gy dose groups. RT-PCR results were consistent with the results of the gene chip, including CDKN2A gene, TNFRSF19, RNASE7 and EGFR3, Real- Time PCR confirmed that TNFRSF19 expression decreased after irradiation. Conclusion Different doses of X-ray irradiation affect the expression of many functional genes in AHH-1 cells, and the differentially expressed genes involved in different biological processes after irradiation.