P08.08 Modulation of CD30 receptor shedding in the presence of EDTA on K562 cells pretreated with TNF

Background The TNF/TNF receptor system plays an important role in signal transduction for the induction of apoptosis and necrosis. TNF receptors are a complex group of proteins consisting of several families of receptors and are activated by the cytokine TNFα. In this work, the role of EDTA as an inhibitor of the TACE signaling pathway on the expression of the surface CD30 antigen on K-562 cells after apoptosis induction was investigated.Material and Method. K562 cells were treated with and without the presence of the cytokine TNF-α at a concentration of 1000 pg/ml for 24 h with and without the presence of EDTA in an incubator with 5% CO2. After treatment, the expression of the CD30 molecule was examined on a BD, Fax verse (USA) flow cytometer, the degree of cell death was examined using LDH release, while CD30 in the supernatant of cell cultures was determined using the Western blot technique.Results The results showed that TNF-a led to a statistically significant decrease in the expression of the CD30 molecule on the membrane of K562 cells (p < 0.05, Mann Whitey, U-test), to a statistically significant increase in LDH inducing cell death. Examination of the expression of CD30 on the membrane of K562 cells after the addition of EDTA, which is an inhibitor of the TACE enzyme responsible for receptor cleavage, resulted in a partially reversible decrease in expression and a slight increase in the value by 10% compared to TNF-α without the presence of EDTA.Conclusion It has been shown that EDTA as an inhibitor of the enzyme that blocks signal transmission through the TNF family of receptors can reduce the effects on cells, but the results are dose-dependent and further studies are needed to show the blockade depending on the receptor in order to achieve a stronger effect. Further studies should be directed to the presence of parts of the molecule in the supernatant of cell cultures in order to prove parts of extracellular receptor fragments.