赖氨酸甲基转移酶PR-Set7及S-腺苷甲硫氨酸在亚砷酸钠调控人永生化皮肤角质形成细胞碱基切除修复基因H4K20me1修饰中的作用  

Effects of lysine methyl transferase PR-Set7 and S-adenosylmethionine on sodium arsenite-mediated modification of base excision repair gene H4K20me1 in human immortalized skin keratinocytes

作　　者：王祺[1] 谢琅[1] 丁雪娇[1] 李昌哲[1] 李军[1] WANG Qi;XIE Lang;DING Xue-jiao;LI Chang-zhe;LI Jun(Key Laboratory of Environmental Pollution Monitoring aiul Disease Control,Ministry of Education,School oj Public Health,Guizhou Medical University,Guiyang,Guizhou 550025,China;不详)

机构地区：[1]贵州医科大学环境污染与疾病监控教育部重点实验室,公共卫生学院,贵州贵阳550025

出　　处：《环境与健康杂志》2019年第6期485-490,共6页Journal of Environment and Health

基　　金：国家自然科学基金(81360411);贵州省科技厅基金(黔科合基础[2018]1134).

摘　　要：目的探讨赖氨酸甲基转移酶PR-Set7和S-腺苷甲硫氨酸(SAM)对亚砷酸钠调控人永生化皮肤角质形成细胞(HaCaT细胞)碱基切除修复(BER)基因组蛋白H4第20位赖氨酸一甲基化(H4K20me1)修饰水平的影响。方法体外培养HaCaT细胞,设对照(24 h)组、NaAsO2(10.00μmol/L,24 h)染毒组、赖氨酸甲基转移酶PR-Set7小干扰RNA(siPR-Set7)干扰组(50 nmol/L,siPR-Set7,48 h)、siPR-Set7+NaAsO2组(50 nmol/L siPR-Set7干扰48 h后,10.00μmol/LNaAsO2处理24 h)和SAM+NaAsO2组(SAM处理1 h后10.00μmol/LNaAsO2处理24 h),染色质免疫共沉淀(CHIP)检测N-甲基化嘌呤-DNA-糖基化酶(MPG)、聚腺苷酸二磷酸核糖聚合酶-1(PARP1)、X射线修复交叉互补基因-1(XRCC1)基因启动子区(CHIP1、CHIP2区域)和编码区(CHIP3、CHIP4区域)H4K20me1修饰水平,实时荧光定量PCR(qRT-PCR)检测MPG、PARP1、XRCC1基因mRNA表达水平。结果与对照组比较,NaAsO2染毒组、siPR-Set7组和siPR-Set7+NaAsO2组MPG、PARP1基因启动子CHIP1区及XRCC1基因启动子CHIP2区H4K20me1富集减少,SAM+NaAsO2组MPG、PARP1基因启动子CHIP1和CHIP2区及XRCC1基因启动子CHIP1区H4K20me1富集减少;与NaAsO2染毒组比较,siPR-Set7+NaAsO2组PARP1基因启动子CHIP1区、XRCC1基因启动子CHIP2区H4K20me1富集减少,SAM+NaAsO2组PARP1基因启动子CHIP1区及XRCC1基因启动子CHIP2区H4K20me1富集增加(P均<0.05)。与对照组比较,NaAsO2染毒组、siPR-Set7组和siPR-Set7+NaAsO2组MPG基因编码CHIP4区及PARP1、XRCC1基因编码CHIP3和CHIP4区H4K20me1富集减少,SAM+NaAsO2组MPG基因编码CHIP3和CHIP4区及PARP1、XRCC1基因编码CHIP3区H4K20me1富集减少;与NaAsO2染毒组比较,siPR-Set7+NaAsO2组MPG基因编码CHIP4区及PARP1、XRCC1基因编码CHIP3和CHIP4区H4K20me1富集减少,SAM+NaAsO2组MPG、PARP1、XRCC1基因编码CHIP3和CHIP4区H4K20me1富集增加(P均<0.05)。与对照组比较,NaAsO2染毒组、siPR-Set7组、siPR-Set7+NaAsO2组和SAM+NaAsO2组MPG、XRCC1、PARP1 mRNA表达降低;与NaAsO2染毒组比较,siPR-SObjective To understand the effect of lysine methyltransferase PR-Set7 and S-adenosylmethionine(SAM) on the modification level of lysine methyl transferase(H4K20me1) in human immortalized skin keratinocytes(HaCaT cells) induced by sodium arsenite. Methods HaCaT cells in culture were divided into the control group(24 h),sodium arsenite exposure group(10 μmol/L,24 h),small interfering RNA of PR-Set7 group(50 nmol/L siPR-Set7,48 h),siPR-Set7+NaAsO2 group(50 nmol/L siPR-Set7 interference 48 h,10 μmol/L sodium arsenite exposure for 24 h) and SAM+NaAsO2 group(SAM,1 h;10 μmol/L sodium arsenite exposure for 24 h). The modification level of H4 K20 me1 in promoter region(CHIP1,CHIP2 region) and coding region(CHIP3,CHIP4 region) of N-methylated purine-DNA-glycosylase(MPG),polyadenylate diphosphate ribose polymerase-1(PARP1),X-ray repair cross-complementary gene-1(XRCC1) were measured by chromatin immunoprecipitation(CHIP). The expression levels of MPG,PARP1 and XRCC1 mRNA were measured by qRT-PCR. Results Compared with the control group,the levels of H4 K20 me1 in CHIP1 region of MPG,PARP1 promoter and CHIP2 region of XRCC1 promoter decreased in NaAsO2 exposure group,siPR-Set7 interference group and siPR-Set7+NaAsO2 group,while those in CHIP1 and CHIP2 regions of MPG and PARP1 promoter and CHIP1 region of XRCC1 promoter decreased in SAM+NaAsO2 group;Compared with NaAsO2 exposure group,the levels of H4 K20 me1 decreased in CHIP1 region of PARP1 gene promoter and CHIP2 region of XRCC1 gene promoter in SiPR-Set7+NaAsO2 group,increased in CHIP1 region of PARP1 gene promoter and CHIP2 region of XRCC1 gene promoter in SAM+NaAsO2 group(all P<0.05). Compared with the control group,the levels of H4 K20 me1 in CHIP4 region of MPG gene coding and CHIP3 and CHIP4 regions of PARP1,XRCC1 gene coding decreased in NaAsO2 exposure group,siPR-Set7 interference group and siPR-Set7+NaAsO2 group,while those in CHIP3 and CHIP4 region of MPG gene coding and CHIP3 region of PARP1 and XRCC1 gene coding decreased in SAM+NaAsO2 group;Compared with

关 键 词：亚砷酸钠 人永生化皮肤角质形成细胞 H4K20甲基化 赖氨酸甲基转移酶PR-Set7 S-腺苷甲硫氨酸 碱基切除修复 

分 类 号：R994.6[医药卫生—毒理学]