16th Horizons in Molecular Biology Göttingen-Germany, Göttingen, Almanya, 9 - 12 Eylül 2019, cilt.1, ss.127
Many cells and tissue express a mixture of cholinergic receptors. Different studies have suggested that nonneuronal acetylcholine and cholinergic agonists alter cell growth and proliferation of T and B lymphocytes. Costa et al. demonstrated that acetylcholine released from T-lymphocytes acts via the M3 acetylcholine muscarinic receptor (mAChR) to trigger signaling pathways. Muscarinic cholinergic receptors activate stimulatory growth mechanisms in megakaryocytic differentiated K562 leukemia cells.
In this study, we investigated the levels of caspase 3,8 and 9 protein expression and caspase activity in megakaryocytic differentiated K562 leukemia cells. And we also investigated the effects of muscarinic agonist effects on apoptosis megakaryocytic differentiated K562 leukemia cells.
Megakaryocytic differentiation was induced by applying phorbol 12-myristate 13-acetate (PMA) (Calbiochem) at the concentration of 10 nM. Anti-Caspase 3,8 and 9 antibodies were purchased from Santa Cruz, Inc. .). For western blot assay, Proteins were electrotransferred to nitrocellulose membranes which were exposed to primary antibodies and then to alkaline phosphatase-conjugated secondary antibodies. The antibody–antigen complex was detected with NBT/BCIP. Caspase activity detected by Elisa technique.
We detected caspase 3,8 and 9 activity and expression in megacaryocytic differentiated K562 leukemia cells. Especially muscarinic receptor agonist and antagonist decrease caspase 3 and caspase 9 expression. These results suggest that CCh may be modulates megakaryocytıc dıfferentiated K562 leukemic cells apoptosis through muscarinic acetylcholine receptors