聚腺苷二磷酸核糖聚合酶1在苯并（a）芘诱导人支气管上皮细胞DNA甲基化改变中的作用  

Role of poly （ADP-ribose） polymerase 1 on DNA methylation variation induced by B （a）P in human bronchial epithelial cell

作　　者：陶功华[1] 龚春梅[1] 杨淋清[1] 刘庆成[1] 刘建东[1] 吴德生[1] 胡辛楠[1] 黄海燕[1] 刘建军[1] 柯跃斌[1] 庄志雄[1] 

机构地区：[1]深圳市疾病预防控制中心毒理研究室,518020

出　　处：《中华预防医学杂志》2011年第5期410-415,共6页Chinese Journal of Preventive Medicine

基　　金：基金项目：国家自然科学基金（30571592,30700673,81072323）;国家重点基础研究发展计划（2002CB512903）

摘　　要：目的 观察苯并（a）芘[benzo（a）pyrene,B（a）P]诱导体外细胞DNA甲基化水平改变,探讨聚腺苷二磷酸核糖聚合酶1[poly（ADP-ribose）polymerase 1,PARP1]在该过程中的作用.方法 以1.0、2.0、5.0、10.0、15.0、30.0 μmol/L浓度B（a）P分别处理人支气管上皮细胞（16HBE）及其PARP1缺陷细胞（16HBE-shPARP1）72 h.采用免疫荧光和高效毛细管电泳检测其基因组DNA整体甲基化水平改变,同时动态监测PARP1和DNA甲基转移酶1（DNA methyltransferases 1,DNMT1）表达的变化.结果 16HBE和16HBE-shPARP1细胞基因组整体甲基化百分比（mCpG%）分别为（4.04±0.08）%和（9.69±0.50）%.经5-氮杂脱氧胞苷（DAC）处理72 h后,mCpG%值分别下降为（3.15±0.14）%、（6.07±0.54）%.经B（a）P染毒72 h后,16HBE细胞基因组mCpG%值[B（a）P浓度由低到高]分别为（5.10±0.13）、（4.25±0.10）、（3.91±0.10）、（4.23±0.27）、（3.70±0.15）、（3.08±0.07）;16HBE-shPARP1细胞基因组mCpG%值（浓度由低到高）分别为（10.63±0.60）、（13.08±0.68）、（9.75±0.55）、（7.32±0.67）、（6.90±0.49）、（6.27±0.21）.两种细胞不同处理组间mCpG%差异有统计学意义（F值分别为61.67、60.91,P值均＜0.01）.16HBE细胞各剂量组[B（a）P浓度由低到高]PARP1基因mRNA相对表达水平分别为对照组的141.0%、158.0%、167.0%、239.0%、149.0%、82.9%,差异均有统计学意义（t值分别为11.45、17.32、32.24、33.44、20.21、9.87,P值均＜0.01）;16HBE-shPARP1细胞各剂量组（浓度由低到高）PARP1基因mRNA相对表达水平分别为对照组的169.0%、217.0%、259.0%、323.0%、321.0%、256.0%,差异有统计学意义（t值分别为9.06、15.92、22.68、26.23、37.19、21.15,P值均＜0.01）.当B（a）P染毒剂量达5.0 μmol/L后,16HBE细胞各剂量组（浓度由低到高）DNMT1基因mRNA相对表达水平分别为对照组的125.0%、162.0%、275.0%、233.0%,差异有统计学意义（t值分别为12.74、24.92、55.1Objective To investigate DNA methylation variation in human cells induces by B （a）P, and to explore the role of PARP1 during this process. Methods The changes of DNA methylation of 16HBE and its PARP1-deficient cells exposed to B （ a ） P （ 1.0, 2. 0, 5.0, 10. 0, 15.0, 30. 0 μ mol/L ） were investigated by immunofluorescence and high performance capillary electrophoresis, and simultaneously, the expression level of PARP1 and DNMT1 were monitored dynamically. Results The percentage of methylated DNA of overall genome （ mCpG% ） in 16HBE and 16HBE-shPARP1 cells were separately （4. 04 ±0. 08） %and （9. 69 ±0. 50）%. After being treated by 5-DAC for 72 hours,mCpG% decreased to （3.15 ±0. 14）%and （6. 07 ± 0. 54 ） %. After both being exposed to B （a） P for 72 hours, the mCpG% in 16HBE group （ ascending rank ） were separately （ 5. 10 ± 0. 13 ）, （ 4. 25 ± 0. 10 ）, （ 3.91 ± 0. 10 ）, （ 4. 23 ± 0. 27 ）,（3.70 ± 0. 15 ）, （ 3.08 ± 0. 07 ）; while the figures in 16HBE-shPARP1 group （ ascending rank ） were respectively （10.63 ±0.60）, （13.08 ±0.68）, （9.75 ±0.55）, （7.32 ±0.67）, （6.90 ±0.49） and （6. 27 ±0. 21 ）. The difference of the results was statistically significant （ F values were 61.67 and 60. 91,P〈 0.01 ） . For 16HBE group, expression of PARP1 and DNMT1 were 141.0%, 158.0%, 167.0%,239. 0%, 149. 0% ,82. 9% and 108. 0%, 117.0%, 125.0%, 162. 0% ,275. 0% ,233.0% comparing with the control group, whose difference also has statitical significance （t values were 11.45,17. 32,32. 24,33.44,20.21 and 9. 87 ,P 〈 0. 01 ）. For 16HBE-shPARP1 group, expression of PARP1 and DNMT1 were 169.0% ,217.0%, 259.0%, 323.0% , 321.0% , 256.0% and 86.0% , 135.0% , 151.0% , 180.0%,229. 0%, 186. 0% comparing with the control group,with statitical significance （t values were 9. 06,15. 92,22. 68,26. 23,37. 19 and 21.15, P 〈 0. 01 ）. When the dose of B （a） P reached 5.0 μmol/L, the mRNA expression of DNMTI in 16HBE group （ascendin

关 键 词：苯并芘 DNA甲基化 聚ADP核糖聚合酶类 DNA(胞嘧啶-5-)-甲基转移酶 

分 类 号：R730.2[医药卫生—肿瘤]