Earticle

After somatic cell nuclear transfer (SCNT), epigenetic state of a differentiated donor cell nucleus must be reversed to the embryonic state. Incomplete epigenetic reprogramming is thought to be causing the low cloning efficiency. The present study was carried out to investigate the effects and mechanism of scriptaid, a novel histone deacetylase inhibitor (HDACi), on the in vitro development of porcine SCNT embryos. We found that treating SCNT embryos with 300 or 500 nM scriptaid for 20 h increased the development of embryos to the blastocyst stage and total cell numbers in per blastocyst( p<0.05). Apoptosis in blastocyst was decreased (p<0.05) in the presence of scriptaid. Scriptaid treatment significantly (p<0.05) increased the levels of H3-acK9 and 5-hmc, and the levels of H3-m3K9 and 5-mc were decreased at the pronuclear stage. Expression level of mir-152 was significantly (p<0.05) increased in scriptaid treated embryos. Moreover, treating embryos with 300 nM scriptaid increased the expression level of the genes that play important roles during embryonic development (OCT4 and CDX2). Additionally, mRNA expression of Dnmt1, Cas3 and Bak were significantly decreased and Bcl-xL was increased in scriptaid treated group compared to control. In conclusion, scriptaid improves the developmental capacity, prevent apoptosis through improving the epigenetic reprogramming in porcine SCNT embryos.

Partheno Embryo's research is known to play a very important role in identifying the development of embryonic cells or analyzing the genetic mechanisms of embryonic development, but the information on apoptosis formed during the early stage of development on Partheno Embryo is very little. Therefore, this study analyzed whether the embryonic cell death of unit embryos can be inhibited by adding Scriptaid, one of HDACi, which plays a role in demethylation of histone proteins as a method of regulating the cell cycle in the early embryo development of Partheno Embryo. As a result, the differentiation rate was higher in the group that added Scriptaid and FBS, but the cellular development was higher in the group that added pregnant serum to Scriptaid. As a result of analyzing the expression of the gene through IF and PCR, the group with the addition of gestational serum increased the expression of BCL2 and PCNA, which affects the anti-Casp3 action in cell survival. In addition, it is interpreted that treatment of Scriptaid for 16 hours, rather than 24 h treatment lowers the expression of Casp-3, a representative factor of apoptosis, and also increases embryonic development, thus affecting early embryo development. Therefore, it is concluded that the 16-hour treatment of Scriptaid and the use of gestational serum will inhibit cell death in the early embryonic development and increase the development rate of the embryo.

Histone deacetylation and demethylation are epigenetic mechanisms implicated in cancer. Studies regarding the role of modulation of gene expression utilizing the histone deacetylase inhibitor scriptaid and the demethylating agent 5-azacytidine in HL-60 leukemia cells have been limited. We studied the possibility of recovering epigenetically silenced genes by scriptaid and 5-azacytidine in human leukemia cells by DNA microarray analysis. The first group was leukemia cells that were cultured with 5-azacytidine. The second group was cultured with scriptaid. The other group was cultured with both agents. Two hundred seventy newly developed genes were expressed after the combination of 5-azacytidine and scriptaid. Twenty-nine genes were unchanged after the combination treatment of 5-azacytidine and scriptaid. Among the 270 genes, 13 genes were differed significantly from the control. HPGD, CPA3, CEACAM6, LOC653907, ETS1, RAB37, PMP22, FST, FOXC1, and CCL2 were up-regulated, and IGLL3, IGLL1, and ASS1 were down-regulated. Eleven genes associated with oncogenesis were found among the differentially expressed genes: ETS1, ASCL2, BTG2, BTG1, SLAMF6, CDKN2D, RRAS, RET, GIPC1, MAGEB, and RGL4. We report the results of our leukemia cell microarray profiles after epigenetic combination therapy with the hope that they are the starting point of selectively targeted epigenetic therapy.