Determination of expression of P97/valosin-containing protein (vcp) in bone marrow-derived cells and investigation of the effect of P97/VCP inhibitor on these cells at children with acute lymphoblastic leukemia (all) and healthy donors

Acute lymphoblastic leukemia (ALL) is a malignant disease, which occurs as a result of mutations that occur during differentiation of lymphoid cells in bone marrow. The bone marrow microenvironment has an effect at the onset of ALL. The most important group of cells of this microenvironment is mesenchymal stem cells (MSCs), which have an important role in the secretion of many cytokines. p97/VCP is a member of AAA+ATPase family. It has an essential role in various important pathways such as intracellular ubiquitin proteasome pathway, endoplasmic-reticulum-associated protein degradation, cell cycle, apoptosis, and autophagy. It is shown that p97/VCP is overexpressed in most of the cancer types and inhibitors for this protein have effects as therapeutic agents on proliferating cancer cells. DBeQ (dibenzylquinazoline-2,4-diamine), is a p97/VCP inhibitör that has been shown to have least toxic effects on cells. The aim of this study is to determine and compare the presence of p97/VCP in mononuclear cells (MNC) and MSCs in ALL patients and healthy donors and to investigate the effect of DBeQ (10μM-1 hour) on these cells. For this purpose, the expressions of p97/VCP, p62 and LC3B in the isolated cells were determined by Western blot analysis and immunofluorescence staining. In addition, the effect of DBeQ on apoptosis was investigated by flow cytometry with Annexin V method. As a result, it was shown that p97/VCP expression was higher in the ALL patient cell groups, the expression of p62 and LC3B decreased after DBeQ which has significant apoptotic activity in ALL cell lines. Keywords: Acute lymphoblastic leukemia, mesenchymal stem cells, p97/VCP, DBeQ