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. 2004 Sep 15;10(18):2643-6.
doi: 10.3748/wjg.v10.i18.2643.

Histone acetylation regulates p21WAF1 expression in human colon cancer cell lines

Ying-Xuan Chen  1 , Jing-Yuan Fang, Hong-Yin Zhu, Rong Lu, Zhong-Hua Cheng, De-Kai Qiu
Affiliations

Histone acetylation regulates p21WAF1 expression in human colon cancer cell lines

Ying-Xuan Chen et al. World J Gastroenterol. .

Abstract

Aim: To investigate the effect of histone acetylation on regulation of p21WAF1 gene expression in human colon cancer cell lines.

Methods: Two cell lines, Colo-320 and SW1116 were treated with either trichostatin or sodium butyrate. Expressions of p21WAF1 mRNA and protein were detected by real-time RT-PCR and Western blotting, respectively. Acetylation of two regions of p21WAF1 gene-associated histones and total cellular histones were examined by chromatin immunoprecipitation assay and Western blotting.

Results: Trichostatin or sodium butyrate re-activated p21WAF1 transcription resulted in up-regulated p21WAF1 protein level in colon cancer cell lines. Those effects were accompanied by an accumulation of acetylated histones in total cellular chromatin and p21WAF1 gene-associated region of chromatin.

Conclusion: Histone acetylation regulates p21WAF1 expression in human colon cancer cell lines, Colo-320 and SW1116.

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Figures

Figure 1
Figure 1
Western blotting of acetylated histones H3, H4 and p21WAF1 protein in Colo-320 cells. Lane 1: Control (PBS); lane 2: Control (alcohol); lane 3: TSA 1 μmol/L, 24 h; lane 4: NaBu 5 mmol/L, 24 h.
Figure 2
Figure 2
RT-PCR showed either TSA or sodium butyrate induced overexpression of p21WAF1 mRNA in human colon cancer cell lines Colo-320 and SW1116. Lane 1: Control (PBS); lane 2: Control (alcohol); lane 3: TSA 1 μmol/L, 24 h; lane 4: NaBu 5 mmol/L, 24 h.
Figure 3
Figure 3
HDAC inhibitors induced accumulation of acety-lated histones H3 and H4 in chromatin associated with p21WAF1 gene. A: PCR products for the 1st set primer of p21WAF1; B: PCR products for the 2nd set primer of p21WAF1; C: The figure in A and B was scanned and quantified by using IMAGE analysis software. The ratio between input DNA and precipitated DNA was calculated for each treatment and primer set. The increase after treatment with either TSA or sodium butyrate was calcu-lated from indicated ratios. PCR products of the 2 nd primer were not visualized on agarose gel before treatment with HDAC inhibitor, so it could not be calculated. Input DNA, lanes 1-4: Soluble chromatin was immunoprecipitated with antiacetylated histone H3 antibody; lanes 5-8: Soluble chro-matin was immunoprecipitated with antiacetylated histone H4 antibody; lanes 9-12: Normal rabbit serum as negative control; lanes 13-16, 1, 5, 9, 13: PBS control; lanes 2, 6, 10, 14: Alcohol control; lanes 3, 7, 11, 15: TSA 1 μmol/L, 24 h; lanes 4, 8, 12, 16: NaBu 5 mmol/L, 24 h.
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