[Effects of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells: a study using chromatin immunoprecipitation].
OBJECTIVE
To develop a real-time PCR-based chromatin immunoprecipitation (ChIP) assay for determining the effect of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells.


METHODS
K562 cells were grown in the presence or absence of 0.5 mmol/L sodium butyrate for 48 h, and 1=10(7) cells per group were used for real-time PCR-based ChIP with anti-acetylated histone H3 or H4 antibodies. The levels of acetylated histone H3 and H4 (acH3 and acH4) in Ggamma- and Agamma-globin gene promoter regions were measured.


RESULTS
In the K562 cells with sodium butyrate treatment or without any treatment, the levels of acH3 or acH4 in Ggamma- or Agamma-globin gene promoter were higher than that in the necdin gene (negative control). Compared with the untreated K562 cells, the cells treated with 0.5 mmol/L sodium butyrate showed a 3.1-fold or 2.6-fold increase in acH3 or acH4 in Ggamma-globin gene promoter region, with also a 3.7-fold or 3.2-fold increase in acH3 or acH4 in Agamma-globin gene promoter region, respectively (P<0.01).


CONCLUSIONS
We have successfully developed a real-time PCR-based ChIP assay for analyzing the acetylation of histone H3 and H4 in gamma-globin gene promoter regions. Our results support the role of sodium butyrate in increasing the level of acetylated histone in gamma-globin gene promoter regions.
Chen
Jian-feng
JF
Department of Neonatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. adjfchen@yahoo.com.cn
Qian
Xin-hua
XH
Zhao
Dan-hua
DH
Qian
Xin-lai
XL
D000107:Acetylation; D002087:Butyrates / Q000494:pharmacology*; D047369:Chromatin Immunoprecipitation / Q000379:methods; D006657:Histones / Q000737:chemistry*; D006801:Humans; D020014:K562 Cells; D011401:Promoter Regions, Genetic / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D055545:gamma-Globins / Q000235:genetics*
D002087:Butyrates; D006657:Histones; D055545:gamma-Globins