J Korean Surg Soc.  2009 May;76(5):279-284. 10.4174/jkss.2009.76.5.279.

The Combination Effect of Sodium Butyrate, 5-aza-2'-deoxycytidine on the Tumor Suppressive Activity in RKO Colorectal Cancer and MCF-7 Breast Cancer Cell Lines

• Cho HJ
• Park SC
• Kim KW
• Kang WK
• Cho HM
• Kim JS
• Kim YA
• An CH

• KMID: 2096547
• DOI: http://doi.org/10.4174/jkss.2009.76.5.279

Abstract

PURPOSE
It is known that DNA methylation is associated with histone acetylation status in regulation of gene expression. In this study, we investigate the effect of demethylating agents and histone deacetylase (HDAC) inhibitor on the tumor suppression and the combined effect of two agents according to methylation status in human colon and breast cancer cell lines. METHODS: In this study, the RKO colorectal cancer cell line, MCF-7 breast cancer cell lines were considered. For each cell line, we used HDAC inhibitor sodium butyrate (SB), demethylating agent 5-aza-2'-deoxycytidine (5-aza-DC) and a combination of both agents. We estimated the percentage of cell survival using the XTT method and experimented with the augmentative effects of both agents. RESULTS: In RKO cell line in which most of the genes are methylated, 74% of cell survival was shown for 5-aza-DC treatment and 83% of cell survival for SB treatment. In MCF-7 cell line that approximately half of the genes are methylated, 82% cell survival was shown for 5-aza-DC treatment and 63% cell survival for SB treatment. We observed that the survival fraction is lower after the combined treatment of 5-aza-DC and SB than that of 5-aza-DC or SB alone in both RKO (53%) and MCF-7 (49%) cell lines (P<0.001). CONCLUSION: For highly methylated genes, 5-aza-DC is more effective on the tumor suppression than SB. On the other hand, if the methylation of the promoter region is at low density, SB is noted to be more effective than 5-aza-DC. Furthermore, the combined treatment of 5-aza-DC and SB is more effective than using each agent alone.

Keyword

Epigenetics; Histone deacetylase inhibitor; Demethylating agent

MeSH Terms

Acetylation
Azacitidine
Breast
Breast Neoplasms
Butyrates
Cell Line
Cell Survival
Colon
Colorectal Neoplasms
DNA Methylation
Epigenomics
Gene Expression Regulation
Hand
Histone Deacetylase Inhibitors
Histone Deacetylases
Histones
Humans
MCF-7 Cells
Methylation
Promoter Regions, Genetic
Sodium
Azacitidine
Butyrates
Histone Deacetylase Inhibitors
Histone Deacetylases
Histones
Sodium

Figure

• 

  Fig. 1 MS-PCR after 5-aza-2'-deoxycytidine (5-aza-DC) treatment. In the control group, most of the genes were methylated, but cell lines treated with aza-DC showed that demethylated bands and methylated bands were slightly decreased.

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  Fig. 2 MS-PCR after sodium butyrate treatment. Compared to the control group, there were almost no changes in methylation status with the addition of sodium butyrate.

• 

  Fig. 3 The effect of 5-azaDC and SB on cell survival.

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