| Abstract [eng] |
Recently, a novel strategy for the treatment of leukemia’s through the modulation of chromatin structure is applicable. In this study, we conducted a detailed analysis of anti-leukemia effects of histone deacetylase (HDAC) inhibitors and their combinations with retinoic acid using human promyelocytic leukemia cell line HL-60. We have shown that HDAC inhibitors - phenyl butyrate and vitamin B3, cause rapid histone H3 and H4 modifications. Further we examined how HDACI and retinoic acid (RA) can modulate gene expression via acetylation and other modifications of histones associated with targeted genes. We performed Chip assay to identify proteins associated with hyperacetylated histone H4. Immunoprecipitated proteins were fractionated by SDS/PAGE and 2DE. Proteomic analysis was performed by using mass spectrometry (MALDI TOF and ESI MS/MS). We identified Sp1 transcriptional activation, DNA methyltransferase, methionine acetyltransferase and other proteins that were associated with modified histones H3 and H4. To evaluate the changes ofp21 gene expression affected by hyperacetylation of histone H4 during HL-60 cell granulocytic differentiation we performed PGR of active chromatin immunoprecipitated with hyperacetylated H4 by using different primers of p21 gene. In this study we have shown that p21 gene expression changes during granulocytic differentiation and depends on inducer. Our results suggest that the chromatin remodeling caused by HDAC inhibitors could be a promising agents in the treatment of human acute promyelocytic leukemia. |
