论文部分内容阅读
目的探讨铜蓝蛋白(Cp)在石英诱导的人胚肺成纤维细胞(HELF)组蛋白修饰改变中的作用以及人第十号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)在其中的影响。方法不同浓度的石英(50、100、200μg/ml)和不同浓度的Cp(10、20、30μg/ml)作用HELF细胞24 h,用蛋白免疫印迹(Western Blot)实验检测蛋白质赖氨酸乙酰化(acety-lys)水平以及组蛋白H2B、H3、H4乙酰化和H3甲基化水平。200μg/ml石英作用于HELF细胞和转染PTEN shRNA的HELF细胞(PT)1 h后,加入30μg/ml Cp作用24 h,用Western blot检测acety-lys水平,组蛋白H2B、H3、H4乙酰化和H3甲基化水平。结果石英暴露可诱导蛋白质acety-lys水平的增高,组蛋白H2B(lys5/12)、H3(lys9/14)、H4(lys12)乙酰化水平增加,组蛋白H3的2位精氨酸的甲基化水平降低。Cp可以逆转石英暴露所致的蛋白质acety-lys水平的增加,组蛋白H3、H4赖氨酸乙酰化水平的增加和组蛋白H3甲基化水平的减少。抑制PTEN的水平后,观察不到Cp对上述现象的逆转。结论 Cp可以逆转石英暴露诱导的组蛋白乙酰化和甲基化改变,PTEN参与了Cp对石英诱导的组蛋白修饰作用的改变。
OBJECTIVE: To investigate the role of ceruloplasmin (Cp) in the alteration of histone-induced histone modifications induced by quartz in human embryo lung fibroblasts (HELF), and to investigate the role of phosphatase and tensin homolog (PTEN) Impact. Methods HELF cells were treated with quartz (50,100,200μg / ml) and different concentrations of Cp (10,20,30μg / ml) for 24 h. Protein lysine acetylation was detected by Western Blot (acety-lys) levels and histone H2B, H3, H4 acetylation and H3 methylation levels. 200μg / ml quartz was applied to HELF cells and PTEN shRNA-transfected HELF cells (PTs) for 1 h, 30 μg / ml Cp was added for 24 h, acety-lys level was detected by Western blot, histone H2B, H3 and H4 acetylated And H3 methylation levels. Results The quartz exposure induced an increase of the acety-lys level, and the histone H2B (lys5 / 12), H3 (lys9 / 14) and H4 Reduce the level of Cp can reverse the increase of acety-lys level caused by quartz exposure, the increase of lysine acetylation level of histone H3, H4 and the decrease of histone H3 methylation level. After suppressing the level of PTEN, no reversal of the above phenomenon by Cp was observed. Conclusion Cp can reverse the histone acetylation and methylation induced by quartz exposure. PTEN is involved in the alteration of quartz-induced histone modification by CpEN.