Investigation of Effect on Epithelial Mesenchymal Transition- EMT Transcription Factors Epigenetic Agents in Acute Myeloid Leukemia

Project no. 23643 · Status: Kapanmış · 2022

Summary

Epithelial mesenchymal transition (EMT) is critical in the development of both myeloid and lymphoid cancers. During the progression of cancer of epithelial origin, activation of the epithelial-mesenchymal transition is associated with metastasis and drug resistance. The EMT process can be defined by identifying the expression changes of EMT-associated transcription factors (TF).  The role of the epithelial-mesenchymal transition in non-epithelial cancer is not clearly known. In this context, investigating epigenetic regulation in terms of EMT transition has become an important issue. Studies in the literature have shown that bromodomain and extraterminal (BET) proteins control EMT transcription factors and are associated with the EMT process, resulting in changes in the expression of epithelial/mesenchymal markers, TF. In the last few years, BET proteins have been identified as critical transcription mediators and have been shown to play important roles, especially in the context of acute leukemia and other hematological cancers, displaying a variety of aberrant transcriptional expression patterns. Acute myeloid leukemia, characterized by low and variable production of healthy hematopoietic cells, is defined as a malignant disease of the bone marrow characterized by clonal expansion and differentiation of myeloid progenitor cells. In this thesis, it was determined that the EMT transcription factor (Vimentin; Snail1; N cadherin; Claudin1; Zeb1; Zo1 E-kadherin; Snail2;Zeb2;Twist1), which is considered as epithelial mesenchymal transition biomarkers, in leukemia cells, by using both mono and combination doses of bromodomain inhibitor PLX51107 and HDAC inhibitor SAHA (vorinostat) It was aimed to determine the expression changes. For this purpose, after culturing the acute myeloid leukemia cell line HL-60 and NCI-BL2171 cells as the control group under appropriate conditions, cell specific mono and synergistic combination IC50 doses determined in accordance with the data obtained from our project coded TGA-2019-20763 were applied to the cells. Afterwards, expression changes at the mRNA level of EMT-associated TFs were determined using Cfx Biorad software.  In line with the data obtained, an expression increase was observed in the VIM gene in mono and combined doses of Plx51107 and Vorinostat. 10-fold at a dose of 6.68 µm Plx51107 in the SNAI1 gene; Approximately 12 layers at a dose of 10 µm Plx51107; Expressive down-regulation was observed 14.74-fold in the vorinosat IC50 dose and finally 17.37-fold in the combination dose. There was also expression decrease in all dose groups of N-cadherin, claudin, Zeb1, E-cadherin, SNAI2 and TWIST1 genes. In the ZO-1 gene, in mono and combined doses of Plx51105 and Vorinostat, max. A 4-fold expression increase was observed. 2.42 for 6.68µm Plx51107 in all dose groups of the ZEB-2 gene, respectively; 2.06 for 10 µm Plx51107; 1.89 for 4.3µm Vorinostat; 1.8-fold increase in expression was seen for the combination dose. As a result, it is thought that with the determination of the expression changes of EMT-associated TFs, it may have a possible therapeutic effect to stop the EMT transition process.

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