The effects of sodium butyrate on the expression of hdac8 gene in ht-29 colorectal cancer cell line

Mahsa Ghiaghi,1 Flora forouzesh,2,* Hamzeh rhimi,3

1. Medical Genomics Research Center, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran.
2. Department of Genetics, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran
3. Department of molecular medicine, Biotechnology Research Center, Pasteur Institute of Iran,Tehran, Iran

Abstract

Introduction

Sodium butyrate (nab) is a potent growth inhibitor and initiates cell differentiation for many cell types in vitro. in this study, we investigated the effects of nab on ht-29 human colorectal cancer cell line and expression of the hdac8 gene. hdac8 is a histone deacetylase that overexpression in cancer cells.

Methods

Ht-29 cell line was maintained in rpmi with 10% fetal bovine serum. 80 x103 cells were seeded into 96-well plates and then treated with various concentrations of nab (from 6.25mm to 200mm). mtt assay was done after 24h, 48h and 72h incubation. then mrna was extracted and cdna was synthesized by yekta tajhiz kit. hdac8 gene expression was investigated by relative quantification real- time pcr method. fold change expression of hdac8 gene after treatment was evaluated by (2-∆∆ct) method.

Results

The results showed that nab inhibits the viability of ht-29 cells. ic50 for ht-29 cells were 50mm, 25mm and 12.5mm after 24h, 48h and 72h respectively. moreover, real- time pcr results showed that in nab treatment cells, the expression of hdac8 gene was significantly decreased compared with the control group (untreated cells) (p< 0.05). in comparison, treated cells were more sensitive to hdaci-induced cell death in dose and time-dependent manner.

Conclusion

Histone deacetylases (hdacs) can act as transcription repressors, due to histone deacetylation, and consequently, hdac inhibitors (hdaci) selectively alter gene transcription. also, they induce apoptosis. therefore, we suggested hdaci can be used as anticancer agents.

Keywords

Hdac inhibitor, sodium butyrate, hdac8 gene, ht-29 cell line