60Coγ射线照射后人肝细胞子代克隆的蛋白质组学研究  

Proteomics study of progeny of normal human liver cells irradiated by 60 Co γ-rays

作　　者：左雅慧[1] 王放[1] 王小莉[1] 李建国[1] 王仲文[2] 童建[3] 

机构地区：[1]中国辐射防护研究院,太原030006 [2]中国原子能科学研究院 [3]苏州大学放射医学与公共卫生学院

出　　处：《中华放射医学与防护杂志》2009年第4期389-392,共4页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的探讨经不同剂量60Coγ射线照射人正常肝细胞后子代克隆的蛋白质表达图谱变化，并对受照射后肝细胞子代的差异蛋白进行质谱鉴定。方法人正常肝细胞7702分别经不同剂量60Coγ射线照射后，存活细胞克隆继续传代培养至40代，提取细胞总蛋白，进行双向电泳测定，凝胶银染和考染显色，用ImageMaster2DPlatium510软件对获得的蛋白图谱加以分析，寻找差异表达的蛋白质。选取差异点，胶内原位酶解后进行MALDI—TOF分析和数据库搜索。结果与对照组相比，各剂量点双向电泳图谱共发现2倍以上差异蛋白质点42个，上调10个，下调32个。成功鉴定出了17个共同差异表达蛋白，这些差异蛋白包括翻译控制肿瘤蛋白、热休克蛋白、氯离子通道蛋白、真核翻译起始因子等。结论不同剂量的照射可导致人肝细胞子代克隆蛋白质图谱的改变，筛选出的差异蛋白可为探讨电离辐射诱发的基因组不稳定性及其分子机制提供实验依据和理论基础。Objective To characterize the differential protein expression in the progeny of human liver cells surviving from ionizing radiation by the proteomic analysis. Methods Two-dimensional electrophoresis gel coupled with mass spectrometry was used to explore the specific protein expression in the progeny of 7702 human liver cells surviving from ionizing radiation. Alterations in expression level of protein spots between the control and the progeny groups were statistically analyzed by ImageMaster 2D Platinum software and mass spectrometry was used to identify the protein spots with significantly altered expression-level. Results The progeny of irradiated cells were derived from human liver cell line exposed to 0, 2, 4, 6 Gy of 60Co γ-irradiation. A total of 42 differentially expressed proteins between the control and the progeny of the irradiated cells groups were screened, of which 17 were identified by matrix assistant laser desorption ion-top off light-mass spectrometry （MALDI-TOFMS） analysis, including 4 up-regulated and 13 down-regulated proteins. Conclusions The differentially expressed proteins profile could he significantly altered in the progeny of irradiated cells. The proteomies approach has the potential to detect the protein changes relevant to radiation-induced genomie instability（RIGI）. Further study of differentially expressed proteins would likely reveal the molecular mechanisms of gene expression in RIGI.

关 键 词：人肝细胞 蛋白质组学 双向电泳 质谱 基因组不稳定性 

分 类 号：R817[医药卫生—影像医学与核医学]