镉处理前后成纤维细胞变异差异分析  

Differential Analysis of Mutation of Fibroblast Cells Before and After Cadmium Treatment

作　　者：纪仲秋[1] 徐建国[2] 何大澄[1] 彭安[1] 管谷茂[3] 铃木信夫[3] 

机构地区：[1]北京师范大学,100875 [2]吉林大学第一医院呼吸内科 [3]日本千叶大学大学院医学研究院

出　　处：《医学研究杂志》2009年第1期55-57,共3页Journal of Medical Research

摘　　要：目的在环境中存在着很多化合物可以引起变异,对于环境的变异原活性的高感度的方法研究是必要的。例如,应用镉对于人体成纤维细胞的致死毒性和变异活性的详细研究报告很少。因此本研究应用紫外线可以检测出的人体细胞变异,用镉处理的重金属镉的变异活性高感度方法能够检测出来,并且活性的特异点进行解明。方法使用培养的人体细胞RSa和派生株。变异原活性的检测应用OuaR(ouabain resistancn)变异原的检出法和癌遗传因子K-ras的盐基置换的变异的differential dot-blot hybridization检测方法。为了变异诱导的频度的检测,进行了MTT(methyl thiazolyl tetrazoliun)法和生存形成率的测定。变异原活性的机制的解明还应用了Western blot法和siRNA方法对热休克蛋白质进行了解析。结果应用镉的变异原活性的盐基置换变异的检出和紫外线程度相同。DNA伤害对应着热休克蛋白(HSP)27蛋白质的细胞内的变化,同样变异频度也出现了高的数值。结论HSP27蛋白高表达的UVr-1细胞比RSa细胞OuaR变异频率要低。用K-ras dot blot方法证明了RSa细胞具有比UVr-1细胞的变异要高。siHSP27细胞观察到OuaR变异的高表达变化以及镉对DNA损伤表现出UVr-1成纤维细胞小于RSa成纤维细胞。ity＇ s method in which the variation of cells by environment can be tested is necessary. For example, the reports of detailed study for the application of cadmium on the human body cell＇ s lethal toxicity and the variation of cells are very few. This research is about a high sensi- tivity method of heavy metal cadmium＇ s variation active processed by cadmium in which the variation of cells can be tested by ultraviolet ray. Methods The study used cultured cells of human being and derivation. The assay of mutagenicity OuaR （Ouabain resistance） varia- tion used picking law and Differential dot - blot of cancer gene K - ras variation the hybridization examination method. In order to test the induction＇ s frequency of cell variation, MTT （ methyl thiazolyl tetrazoliun） and survival efficiency of formation were tested. The clarifing of mutagenicity mechanism used Western blot and the siRNA method to analysis the molecular of heat shock protein. Results Cadmium clarifiing mutagenicity variation detection had the same degree with the method of ultraviolet ray. The DNA was injury corresponding to the HSP27 protein cell＇ s change. The cell variation frequency also presented the high value. Conelusion Human fibroblasts in the role of the heavy metal cadmium showed that HSP27 protein expression in high - UV^r - 1 cells RSa has lowor - frequency variations than OuaR cells. K - ras dot blot method proved that RSa cells variation is higher than the cells UVr - 1 mutation. In siHSP27 cells, we observed Oua^R high variability of expression. Cadmium on DNA damage showns UV^r - 1 fibroblast cell is lower than RSa fibroblasts cell.

关 键 词：镉 变异 癌因子K-ras 热休克蛋白27 

分 类 号：R730.231[医药卫生—肿瘤] R619.6[医药卫生—临床医学]