镍冶炼烟尘对小鼠胚胎成纤维细胞CpG岛甲基化的影响  

Effect of nickel smelting dusts on methylation of CpG islands of mouse embryonic fibroblasts

作　　者：马文翰[1] 王玥[1] 张琳[1] 于翠平[1] 吴永会[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院,黑龙江哈尔滨150081

出　　处：《中国工业医学杂志》2015年第1期3-7,F0003,共6页Chinese Journal of Industrial Medicine

基　　金：国家自然科学基金项目(编号:81072283)

摘　　要：目的探讨镍冶炼烟尘对小鼠胚胎成纤维细胞(NIH/3T3)中O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)以及死亡相关蛋白激酶1(DAPK 1)启动子CpG岛甲基化程度的影响。方法采集某镍冶炼厂的镍冶炼烟尘,玛瑙研钵研磨磨至直径≤5μm的颗粒占95%以上,称取一定量研磨后的镍冶炼烟尘用PBS溶液配制浓度为2 mg/ml的悬浊液。将处于对数生长期的NIH/3T3细胞分别接触浓度为0、25、50、75、100、125、150μg/ml的镍冶炼烟尘悬浊液24 h后,使用亚硫酸氢盐测序法(bisulfite sequencing PCR,BSP)对接触不同浓度镍冶炼烟尘的NIH/3T3细胞进行MGMT和DAPK 1启动子CpG岛甲基化程度检测。结果与阴性对照组相比,接触镍冶炼烟尘后,NIH/3T3细胞的MGMT启动子CpG岛甲基化程度随着受试物浓度的升高,分别升高了0.5%、0.5%、1.0%、1.0%、2.0%、2.5%;DAPK 1启动子CpG岛甲基化程度随着受试物浓度的升高,分别升高了0.4%、0.5%、0.7%、1.0%、1.7%、1.7%。经Cochran-Armitage趋势检验,MGMT启动子CpG岛甲基化程度与接触镍冶炼烟尘浓度之间呈线性变化趋势,甲基化程度随着接触浓度增加而呈增加的趋势,P值分别为0.0274和0.0281。结论接触不同浓度的镍冶炼烟尘后,NIH/3T3细胞的MGMT以及DAPK 1启动子CpG岛甲基化程度均发生不同程度的升高,且启动子CpG岛甲基化程度与接触镍冶炼烟尘浓度之间呈线性变化趋势,甲基化程度随着接触浓度增高呈增加的趋势。Objective The aim of this research was explore the influence of nickel refining dusts on promoter CpG islands methylation in MGMT and DAPK 1 of mouse embryonic fibroblasts （MEF） NIH/3T3. Methods The nickel refining dusts were collected from the nickel smelting workshop in a factory, and grinded with agate mortar, to the diameter of 95% or more of the particles were less than 5 μm. Prepare the suspension of concentration of 2 mg/ml with PBS solution using the grinded nickel dust particles. Put exponential phase NIH/3T3 cells into the suspensions with different concentrations of nickel refining dusts （0, 25, 50, 75, 100, 125, 150 μm/ml respectively） for 24 hours. Then, measure the methylation level of promoter CpG in MGMT and DAPK 1 of NIH/3T3 after exposing nickel refining dusts with bisulfite sequencing PCR. Results The results showed that compared with negative control group, after exposing to nickel refining dusts, promoter CpG methylation of NIH/ 3T3 in MGMT was increased with the concentration of nickel dust, there were 0. 5%, 0. 5%, 1.0%, 1.0%, 2.0%, 2. 5%, respectively ; the promoter CpG island methylation in DAPK 1 also showed increase with the growing concentration of the nickel dusts, they were increased by 0. 4%, 0. 5%, 0. 7%, 1.0%, 1.7%, 1.7%, respectively. Cochran-Armitage trend test showed that promoter CpG island methylation both in MGMT and DAPK 1, all presented a linear trend with the exposed concentration of nickel refining dusts the P values were 0. 0274 and 0. 0281, respectively. Conclusion The results suggested that after exposure to nickel refining dusts, the methylation of promoter CpG island of NIH/3T3 whether in MGMT or DAPK 1, were all increased in a linear trend with the concentration of nickel refining dusts in exposed suspensions.

关 键 词：镍冶炼烟尘 小鼠胚胎成纤维细胞(NIH/3T3) O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT) 死亡相关蛋白激酶1(DAPK 1) 甲基化 

分 类 号：R135.2[医药卫生—劳动卫生] R994.3[医药卫生—公共卫生与预防医学]