苯并(<i>a</i>)芘致人支气管上皮细胞DNA损伤及与组蛋白H2A单泛素化水平的关系

王志武; 贾利娜; 牛侨; 杨瑾

苯并(a)芘致人支气管上皮细胞DNA损伤及与组蛋白H2A单泛素化水平的关系

Correlation between DNA Damage and Expression of Mono-ubiquitinated Histone H2A in 16HBE Cells Exposed to Benzoapyrene

摘要

摘要:
目的通过体外细胞培养途径,探讨苯并(a)芘(benzoapyrene,BaP)作用下人支气管上皮细胞(16HBE)DNA损伤与组蛋白H2A单泛素化表达水平的关系。

方法取对数生长期16HBE细胞,采用2 μmol/L BaP染毒不同时间(0、1、2、4、8、12、24、48 h)进行实验,用碱性单细胞凝胶电泳实验检测细胞DNA损伤水平,并以Olive尾矩值(olivetail moments,OTM)评价DNA损伤程度;用Western-blot法检测组蛋白H2A单泛素化水平。

结果单细胞凝胶电泳结果显示:2μmol/L BaP染毒不同时间时,随着染毒时间的延长,各染毒组细胞DNA损伤程度均明显增高,与对照组相比,差异均有统计学意义(P<0.01);Western-blot结果显示:2 μmol/L BaP染毒不同时间时,与对照组相比,组蛋白H2A的单泛素化水平增高,且在染毒2 h后,差异有统计学意义(P<0.01);回归分析显示,单泛素化H2A的表达水平与DNA损伤程度存在高度相关,决定系数(R²)为0.910,P<0.01。

结论 BaP染毒致16HBE细胞DNA损伤与组蛋白H2A单泛素化水平之间存在密切关系。

Abstract:
Objective To investigate the association between DNA damage and expression of mono-ubiquitinated histone H2A in human bronchial epithelial cells (16HBE) exposed to benzoapyrene (BaP).

Methods The 16HBE cells collected at the logarithmic phase of growth were exposed to BaP at the concentration of 2μmol/L for 0, 1, 2, 4, 8, 12, 24 and 48 h respectively. DNA damage was determined by comet assay and expressed as olive tail moments (OTM). Protein expression was detected by Western-blot.

Results The results of comet assay showed that the OTM values increased with exposure time and a significant in crement was found as compared with the control group (P<0.01). After exposed to 2μmol/L BaP for 2 h, the mono-ubiquitination levels of histone H2A increased significantly when compared with the control group (P<0.01). The regression analysis showed a significant correlation between OTM and the levels of mono-ubiquitinated histone H2A, and the determination coefficient was 0.910 (P<0.01).

Conclusion The findings suggest a close correlation between DNA damage and the levels of mono-ubiquitinated histone H2A in 16HBE cells exposed to BaP.

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