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目的探讨丁酸钠(NaB)对K562细胞Gγ-珠蛋白基因和Aγ-珠蛋白基因启动子区域组蛋白H3磷酸化/乙酰化(ph/acH3)的影响。方法将K562细胞按不同处理方法分为2组,即0.5 mmol.L-1NaB处理48 h的K562细胞组[K562(NaB)组]和K562亲本细胞组(K562组)。采用半定量RT-PCR测定Gγ-珠蛋白和Aγ-珠蛋白mRNA水平,采用基于Real time PCR分析的染色质免疫共沉淀(ChIP)方法分析不同处理细胞Gγ-珠蛋白和Aγ-珠蛋白基因启动子区域ph/acH3水平。结果与K562组细胞比较,K562(NaB)组细胞Gγ-珠蛋白mRNA、Aγ-珠蛋白mRNA的相对水平分别升高1.4倍(t=-149.022,P=0.000)和1.2倍(t=-13.363,P=0.000)。与K562组比较,K562(NaB)组细胞Gγ-珠蛋白和Aγ-珠蛋白基因启动子区ph/acH3水平分别升高了2.9倍(t=-12.833,P=0.006)和3.2倍(t=-10.484,P=0.000)。K562(NaB)组细胞Gγ-珠蛋白和Aγ-珠蛋白基因启动子片段的%In-put值分别比Necdin基因升高10.0倍(P=0.000)、9.5倍(P=0.000);K562组细胞的Gγ-珠蛋白和Aγ-珠蛋白基因启动子片段的%Input值分别比Necdin基因升高3.2倍(P=0.000)、2.7倍(P=0.000)。结论 NaB可促使γ-珠蛋白基因启动子区域组蛋白H3磷酸化及乙酰化,这一作用途径可能是NaB诱导K562细胞γ-珠蛋白基因表达的机制之一。
Objective To investigate the effect of sodium butyrate (NaB) on histone H3 phosphorylation / acetylation (ph / acH3) in Gγ-globin gene and Aγ-globin gene promoter region of K562 cells. Methods K562 cells were divided into two groups according to different treatment methods: K562 (K562 (NaB) group and K562 parental cell group (K562) treated with 0.5 mmol.L-1NaB for 48 h. Gγ-globin and Aγ-globin mRNA levels were determined by semi-quantitative RT-PCR. The Gγ-globin and Aγ-globin genes were analyzed by ChIP assay using Real time PCR analysis Sub-area ph / acH3 level. Results Compared with K562 cells, the relative levels of Gγ-globin mRNA and Aγ-globin mRNA in K562 (NaB) group were increased by 1.4 times (t = -149.022, P = 0.000) and 1.2 times , P = 0.000). Compared with the K562 group, the levels of ph / acH3 in the promoter region of Gγ-globin and Aγ-globin gene of K562 (NaB) group were increased by 2.9 times (t = -12.833, P = 0.006) -10.484, P = 0.000). The% In-put values of Gγ-globin and Aγ-globin gene promoter fragments of K562 (NaB) group were 10.0 times (P = 0.000) and 9.5 times (P = 0.000) higher than those of Necdin gene respectively. The% Input values of the Gγ-globin and Aγ-globin gene promoter fragments were 3.2-fold (P = 0.000) and 2.7-fold (P = 0.000) higher than that of the Necdin gene, respectively. Conclusion NaB can promote phosphorylation and acetylation of histone H3 in the promoter region of γ-globin gene, which may be one of the mechanisms of NaB-induced γ-globin gene expression in K562 cells.