顺铂对A549细胞DNA损伤修复生物标记物表达的影响  

Study on the expression of DNA excision repair biomarkers in cispatin-treated lung cancer cell line

作　　者：王志勇[1] 周晟[1] 吕斌[1] 张霞[1] 郝巧玲[1] 周宜开[1] 

机构地区：[1]华中科技大学同济医学院环境医学研究所,武汉430030

出　　处：《中华预防医学杂志》2003年第2期109-111,共3页Chinese Journal of Preventive Medicine

基　　金：国家自然科学资金重大项目 ( 39990 5 70 )

摘　　要：目的　研究顺铂对肺癌A5 49细胞中切除修复鼠缺陷交叉互补基因 2 (ERCC2 )蛋白、尿嘧啶DNA糖基化酶 (UDG)、细胞增殖核抗原 (PCNA)在肺癌和食管癌组织中表达水平的影响。方法　通过彗星试验、RT PCR和蛋白印记法研究 3种生物标记物在顺铂处理和未处理A5 49细胞之间的表达水平。分为染毒前、染毒 12h、染毒 2 4h、停止染毒 12h、停止染毒 2 4h 5个不同时间段组 ,每组细胞数均为 1× 10 6个 /ml。结果　在低浓度顺铂 (IC2 0 剂量 )作用下 ,染毒 2 4h内DNA损伤程度的变化与作用时间成正比 ,染毒 12h、2 4h尾相 (单位 :mm)分别为 5 0 2± 0 6 8和 7 2 2± 0 5 3,与阴性对照的尾相2 73± 0 2 9比较有明显差异。停止染毒 2 4h尾相为 3 6 4± 0 70 ,与阴性对照比较无明显差异。ERCC2、UDG和PCNA的mRNA水平和蛋白质水平在细胞染毒后均明显升高 ,染毒 2 4h后mRNA水平分别为 0 71± 0 0 8、0 74± 0 0 6和 0 82± 0 0 9,均明显高于阴性对照 (分别为 0 2 8± 0 0 5、0 31± 0 0 5和 0 37± 0 0 6 ) ;蛋白质表达水平分别为 4 37± 0 5 7、5 47± 0 46和 2 2 1± 0 47,均明显高于阴性对照(分别为 2 2 1± 0 47、2 5 4± 0 38和 3 2 1± 0 47)。停止染毒后 3种酶的mRNA分别为 0 31±Objective To study the expression levels of ERCC2, UDG, and PCNA in cisplatin-treated A549 cell line. Method Comet assay, RT-PCR, and western blot were used to study the mRNA and protein expression levels of ERCC2, UDG,and PCNA. Results When treated with IC 20 cisplatin, the DNA damage level increased as the cisplatin treated time increased within 24 h of cisplatin treatment. The tail state 12 h and 24 h after treatment was 5.02±0.68 and 7.22±0.53 respectively,which was significantly higher than those of the controls(2.73±0.29). The tail state 24 h after treatment was not significantil different from that of the controls. The DNA damage level decreased to normal after cisplatin treatment in 24 h (tail state 3.64±0.7). The expression levels of ERCC2, UDG, PCNA protein (4.37±0.57,5.47±0.46,2.21±0.47 respectively) and mRNA (0.71±0.08,0.74±0.06,0.82±0.09) were increased significantly within 24 h exposure and decreased to normal 24 h after cisplatin treatment. The 3 enzymes' mRNA and protein expression increased when treated with cisplatin, but the changes of protein level were slower than those of mRNA levels. Conclusions The DNA repair capability in A549 cells increases after cisplatin treatment. Cisplatin was a positive regulation of ERCC2, UDG, PCNA expression levels, which causes the increase of mRNA, and protein. The positive regulation only works in a short time and returns normal after 24 h of cisplatin treatment.

关 键 词：肺癌 A549细胞 DNA损伤 顺铂 缺陷交叉互补基因2 尿嘧啶DNA糖基化酶 细胞增殖核抗原 

分 类 号：R73-36[医药卫生—肿瘤]