CAJ | 학술논문

目的比较高低剂量X射线照射对基因表达影响的差异性,探讨不同剂量辐射生物效应的作用机制.方法采用包含有45 033个基因的NimbleGen 12×135K芯片分析0.1和5 Gy照射正常人淋巴母细胞AHH-1培养6h后基因表达谱的改变,以差异为2倍以上且两组实验结果一致的标准确定为有效差异表达基因.用real-time PCR和Western blot方法验证了共同表达基因PERP的表达.结果 0.1 Gy照射后,有760个基因表达上调,1222个基因表达下调；5 Gy照射后,上调和下调基因分别是457和744个.在两个剂量照射条件下共同表达上调的基因有55个,同时下调的基因有339个.低剂量组差异表达基因主要参与细胞信号转导、DNA损伤反应等过程,高剂量组主要参与凋亡、细胞增殖分化等过程.real-time PCR和Western blot方法验证了PERP的表达与芯片结果相一致,均表达下调.结论高低剂量照射后基因表达改变存在质和量的差异,可更好地阐释高低剂量电离辐射生物学效应的作用机制.
목적 비교고저제량X사선조사대기인표체영향적차이성,탐토불동제량복사생물효응적작용궤제.방법 채용포함유45 033개기인적NimbleGen 12×135K심편분석0.1화5 Gy조사정상인림파모세포AHH-1배양6h후기인표체보적개변,이차이위2배이상차량조실험결과일치적표준학정위유효차이표체기인.용real-time PCR화Western blot방법험증료공동표체기인PERP적표체.결과 0.1 Gy조사후,유760개기인표체상조,1222개기인표체하조；5 Gy조사후,상조화하조기인분별시457화744개.재량개제량조사조건하공동표체상조적기인유55개,동시하조적기인유339개.저제량조차이표체기인주요삼여세포신호전도、DNA손상반응등과정,고제량조주요삼여조망、세포증식분화등과정.real-time PCR화Western blot방법험증료PERP적표체여심편결과상일치,균표체하조.결론 고저제량조사후기인표체개변존재질화량적차이,가경호지천석고저제량전리복사생물학효응적작용궤제.
Objective To compare the gene expression difference between 0.1 and 5 Gy X-ray irradiated cells,and to explore its possible mechanism.Methods A cDNA microarray corresponding to 45033 human genes was used to analyze the transcriptional profiles of normal human lymphoblastoid AHH-1 cells at 4 h after 0.1 or 5 Gy irradiation.The genes with a fold change ≥ 2.0 were identified as the differentially expressed genes.real-lime PCR and Western blot were used to confirm the expression of PERP.Results The microarray assay showed that there were 760 up-regulated genes and 1222 down-regulated genes in the cells at 0.1 Gy,while there were 744 genes down-regulated and 457 genes up-regulated in the cells at 5 Gy.In addition,55 genes were commonly up-regulated and 339 genes commonly down-regulated at 0.1 and 5 Gy.The predominant biological processes of the differential genes responding to low-dose radiation include cell-cell signaling transduction and DNA damage response,and the altered genes after 5 Gy irradiation were related to cell proliferation,differentiation,and apoptosis.Moreover,the expression of PERP gene was down regulated,which was consistent with the data of microarrey assay.Conclusions The quantitative and qualitative differences in the gene expressions may contribute to the diversed biological effects induced by low or high doses of ionizina radiation.