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丁雪娇, 谢琅, 陈丽, 马璐, 张爱华, 李军. 亚砷酸钠对人皮肤角质形成细胞BER相关基因H4K20me1修饰水平及其mRNA转录水平影响[J]. 中国公共卫生, 2018, 34(9): 1246-1249. DOI: 10.11847/zgggws1117420
引用本文: 丁雪娇, 谢琅, 陈丽, 马璐, 张爱华, 李军. 亚砷酸钠对人皮肤角质形成细胞BER相关基因H4K20me1修饰水平及其mRNA转录水平影响[J]. 中国公共卫生, 2018, 34(9): 1246-1249. DOI: 10.11847/zgggws1117420
Xue-jiao DING, Lang XIE, Li CHEN, . Effect of sodium arsenite on transcription of base excision repair-related genes and modification of H4K20me1 in HaCaT cells[J]. Chinese Journal of Public Health, 2018, 34(9): 1246-1249. DOI: 10.11847/zgggws1117420
Citation: Xue-jiao DING, Lang XIE, Li CHEN, . Effect of sodium arsenite on transcription of base excision repair-related genes and modification of H4K20me1 in HaCaT cells[J]. Chinese Journal of Public Health, 2018, 34(9): 1246-1249. DOI: 10.11847/zgggws1117420

亚砷酸钠对人皮肤角质形成细胞BER相关基因H4K20me1修饰水平及其mRNA转录水平影响

Effect of sodium arsenite on transcription of base excision repair-related genes and modification of H4K20me1 in HaCaT cells

  • 摘要:
      目的  了解不同剂量亚砷酸钠(NaAsO2)对人永生化皮肤角质形成细胞(HaCaT细胞)碱基切除修复(BER)基因聚腺苷酸二磷酸核糖聚合酶 – 1(PARP1)、N – 甲基化嘌呤 – DNA – 糖基化酶(MPG)、X射线修复交叉互补基因 – 1(XRCC1)启动子区和编码区组蛋白H4第20位赖氨酸一甲基化(H4K20me1)修饰水平及其mRNA转录水平的影响。
      方法  以0.00、2.50、5.00、10.00 μmol/L的NaAsO2处理HaCaT细胞24 h。应用实时荧光定量PCR(qRT-PCR)检测MPG、PARP1、XRCC1基因mRNA表达水平;定量染色质免疫沉淀技术(CHIP-qPCR)检测MPG、PARP1、XRCC1基因启动子区(CHIP1、CHIP2区域)和编码区(CHIP3、CHIP4区域)H4K20me1修饰水平。
      结果  (1)MPG、PARP1、XRCC1基因mRNA表达水平检测结果:2.5 μmol/L染砷组MPG、PARP1、XRCC1基因mRNA表达水平与对照组比较,差异无统计学意义(P > 0.05),5.00、10.00 μmol/L组mRNA表达水平随染毒剂量的增加而降低,与对照组比较,各差异有统计学意义(P < 0.05)。(2)MPG、PARP1、XRCC1基因启动子区和编码区组蛋白H4K20me1修饰水平检测结果:基因启动子区,不同染砷组HaCaT细胞MPG基因启动子CHIP1、CHIP2区和PARP1基因启动子CHIP2区未观察到H4K20me1的富集规律,与对照组相比各差异无统计学意义(P > 0.05),5.00、10.00 μmol/L染毒组HaCaT细胞XRCC1基因启动子CHIP1、CHIP2区和10.00 μmol/L染毒组PARP1基因启动子CHIP1区H4K20me1的富集与对照组相比明显减少,各差异有统计学意义(P < 0.05);基因编码区,5.00、10.00 μmol/L染毒组HaCaT细胞MPG、PARP1、XRCC1基因编码CHIP3、CHIP4区H4K20me1的富集与对照组相比明显减少,差异均有统计学意义(P < 0.05)。
      结论  NaAsO2可通过抑制HaCaT细胞PARP1、XRCC1基因启动子区和MPG、PARP1、XRCC1基因编码区H4K20me1的表达水平,使MPG、PARP1、XRCC1基因mRNA转录水平下降。

     

    Abstract:
      Objective  To investigate the effects of different doses of sodium arsenite (NaAsO2) on mRNA transcription of N-methylpurine-DNA glycosylase (MPG), poly (adenosine diphosphate-ribose) polymerase 1 (PARP1), and x-ray repair cross-complementing 1 (XRCC1) gene and its modification on H4K20me1 of promoter region and coding region in human keratinocyte (HaCaT) cells.
      Methods  HaCaT cells were treated with 2.50, 5.00, 10.00 μmol/L NaAsO2 for 24 hours and a blank control group without NaAsO2 treatment was established. Quantitative real-time polymerase chain reaction was used to detect mRNA expressions of MPG, PARP1, and XRCC1 gene. Quantitative chromatin immuno-precipitation was adopted to determine the level of H4K20me1 modifications in the promoter region (CHIP1 and CHIP2) and coding region (CHIP3 and CHIP4) of MPG, PARP1, XRCC1 gene.
      Results  The mRNA levels of MPG, PARP1, and XRCC1 gene were not significantly different from those of the control group at the NaAsO2 dosage of 2.5 μmol/L (P > 0.05) but the levels gradually decreased significantly at the dosages of 5.00 and 10.00 μmol/L (both P < 0.05). Compared with those of the control group, the enrichments of H4K20me1 in CHIP1 and CHIP2 regions of MPG promoter gene and CHIP2 regions of promoter PARP1 gene in all NaAsO2 dosage groups were not significantly different (P > 0.05); while, the enrichments of H4K20me1 in CHIP1 and CHIP2 regions of XRCC1 promoter gene and CHIP1 regions of PARP1 promoter gene in 5.00 and 10.00 μmol/L dosage groups were reduced significantly (P < 0.05 for all); the enrichment of H4K20me1 in CHIP3 and CHIP4 regions of MPG, PARP1, XRCC1 coding gene were also reduced significantly for the 5.00 and 10.00 μmol/L dosage groups (all P < 0.05).
      Conclusion  NaAsO2 could decrease the mRNA transcription level of MPG, PARP1 and XRCC1 genes by inhibiting the level of H4K20me1 of PARP1, XRCC1 gene promoter region and MPG, PARP1, XRCC1 gene coding region in HaCaT cells.

     

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