Effect of trichostatin A on MHC, histone deacetilaze and DNA methylteransferase gene expression in somatic cell nucleus transfer stem cells

Background: Embryonic stem cells (ESCs) can differentiate into whole cell of the body, for this reason ESCs can be used for therapeutic goals in regenerative medicine. Somatic cell nuclear transfer (SCNT) is an alternative approach for produce ESCs without fertilization. Trichostatin A (TSA) is a HDACi that increases the histones acetylation and thus enhances expression transcriptionally silent allele of imprinted genes. TSA might promote the reprogramming process and improve cloned embryo development. The aim of this study was determination of the effect of Trichostatin A on MHC, histone deacetilaze and DNA methylteransferase gene expression in somatic cell nucleus transfer stem cells.Methods: In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were activated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were established. We selected 15 genes which consisted of MHC- (Qa-1, Qa-2, CIITA, H2db, H2dd , H2KB, H2KD, H2-IE-bb, H2-IE-bd), DNA DNA methylation - (Dnmt-1, Dnmt3a, Dnmt3b), Histone deacetylase- (Hdac1, Hdac2, Hdac3), genes in embryonic stem cell derived from blastocyst with in vitro treated with and without TSA blastocysts (group 1 and 2 )and in vivo blastocysts (control group). For Measurement of gene expression real-time PCR (RT-PCR) was used. All statistical analysis was performed using SPSS 11.5 software with p < 0.05 indicating significance.Result: The blastocyst formation rate of the SCNT embryos treated with 100 nM TSA was higher than that of untreated embryos and control group. Stem cells in group 1 displayed up-regulated expressions of genes including Qa-2and H2-IE-Bd (PConclusion: TSA has positive effect on growth and development of SCNT embryos but TSA leads to abnormal changes in MHC, Dnmt, HDAC gene expression in ESCs cell lines.

Keywords:

Epigenetics modification , Trichostatin A , Somatic cell nuclear transfer