组蛋白H4K20甲基化修饰对砷致HaCaT细胞DNA双键断裂损伤的影响

发布时间：2019-02-14 06:56

【摘要】：[目的]观察亚砷酸钠(NaAsO_2)对人永生化皮肤角质形成细胞(HaCaT细胞)DNA双链断裂损伤、组蛋白H4第20位赖氨酸一、二甲基化(H4K20me1、H4K20me2)修饰水平的影响,探讨H4K20me1、H4K20me2修饰在砷致DNA双键断裂损伤中的作用。[方法]体外常规培养HaCaT细胞,以0.00、1.25、2.50、5.00、10.00μmol/L NaAsO_2连续处理HaCaT细胞24 h,10.00μmol/L NaAsO_2处理HaCaT细胞0、6、12、24 h,其中以0.00μmol/L浓度组和0 h为空白对照组。采用中性单细胞凝胶电泳法检测各组HaCaT细胞DNA双链断裂损伤水平(尾部DNA百分含量、Olive尾矩);免疫印迹法检测各组H4K20me1、H4K20me2蛋白表达水平。[结果]HaCaT细胞染砷24 h后,DNA双链断裂程度在5.00、10.00μmol/L浓度组高于对照组(P0.05);H4K20me1/me2蛋白表达水平在2.50、5.00、10.00μmol/L浓度组低于对照组(P0.05)。10.00μmol/L NaAsO_2处理HaCaT细胞0、6、12和24 h后,DNA双链断裂程度在12、24 h染砷组高于对照组(P0.05),H4K20me1、H4K20me2蛋白表达水平在6、12、24 h较对照组降低(P0.05)。尾部DNA百分含量与H4K20me1、H4K20me2修饰水平呈负相关(r=-0.955、-0.855,均P0.05),Olive尾矩与二者亦呈负相关(r=-0.940、-0.841,均P0.05)。[结论]砷可导致HaCaT细胞DNA双键断裂损伤和H4K20me1、H4K20me2表达改变,提示砷所致DNA损伤可能与组蛋白H4K20甲基化修饰有关。
[Abstract]:[objective] to observe the effect of sodium arsenite (NaAsO_2) on DNA double strand break damage of human immortalized skin keratinocytes (HaCaT cells) and the modification level of H4K20me1H4K20me1 (H4K20me1H4K20me2). Effect of H4K20me2 modification on DNA double bond fracture damage induced by arsenic. [methods] HaCaT cells were cultured routinely in vitro. HaCaT cells were treated with 0.001. 25 ~ 2. 50 渭 mol/L NaAsO_2 and 10. 00 渭 mol/L NaAsO_2 for 24 h, respectively. HaCaT cells were treated with 10. 00 渭 mol/L NaAsO_2 for 24 h. The concentration of 0.00 渭 mol/L and 0 h were taken as the blank control group. The level of DNA double strand break (DNA content in tail, tail moment of Olive) and the expression of H4K20me1H4K20me2 protein were detected by neutrophil gel electrophoresis (neutrophilic single cell gel electrophoresis) and Western blot. [results] after arsenic exposure of HaCaT cells for 24 h, the degree of DNA double strand breaks in the 5.00 渭 mol/L group was higher than that in the control group (P0.05). The expression level of H4K20me1/me2 protein in the 2.50 渭 mol/L NaAsO_2 group was lower than that in the control group (P0.05). The HaCaT cells were treated with 10.00 渭 mol/L NaAsO_2 for 12 and 24 hours. The degree of double strand break of DNA in arsenic exposed group was higher than that in control group (P0.05), and the protein expression level of H4K20me1H4K20me2 was lower than that of control group (P0.05). There was a negative correlation between tail DNA content and H4K20me2 modification level (r ~ (-0.955) ~ (-0.855), P 0.05) and tail moment of), Olive (r ~ (-0.940) ~ (-0.841), P _ (0.05). [conclusion] arsenic can cause DNA double bond breaking damage and H4K20me2 expression change in HaCaT cells, suggesting that DNA damage induced by arsenic may be related to H4K20 methylation modification of histone.
【作者单位】：贵州医科大学公共卫生学院环境污染与疾病监控省部共建教育部重点实验室;
【基金】：国家自然科学基金(编号:81360411);国家自然科学基金重点项目(编号:81430077)
【分类号】：R114

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