The effect of combination of cholinergic agonist and antagonist onapoptosis in megacaryocytic differentiated K562 leukemia cells.


Cabadak H. , Aydın B.

16th Horizons in Molecular Biology Göttingen-Germany, Göttingen, Almanya, 9 - 12 Eylül 2019, cilt.1, ss.127

  • Yayın Türü: Bildiri / Özet Bildiri
  • Cilt numarası: 1
  • Basıldığı Şehir: Göttingen
  • Basıldığı Ülke: Almanya
  • Sayfa Sayıları: ss.127

Özet

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