Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB088

Poster Communications

Cholinergic receptor agonist, ,carbachol, induces M1 and M4 muscarinic receptors expression in megakaryocytic differentiated K562 cells

H. Cabadak1, B. Aydin1

1. Biophysics, Marmara University School of Medicine, Istanbul, Turkey.

  • The effect of CCh, atropine on the expression of M1 (a);M4 (b)muscarinic receptors in megakaryocytic differentiated K562 cells, respectively.*(P < 0.05).

Experimental studies show that muscarinic acetylcholine receptors are potential pharmacological targets for the treatment of cancer (1). Muscarinic receptors have been detected in different cells and tissue (2). Phorbol 12-myristate13-acetate (PMA) causes megakaryocytic differentiation of K562 leukemia cells (3). Previously we showed that cholinergic agonist, carbachol (CCh) increased proliferation of megakaryocytic differentiated K562 cells proliferation in 24 and 48 hours (4). Preclinical studies suggested that cholinergic agents can be useful in cancer treatment prostate cancer (5). In order to detect the effects of agonist stimulation of M1, M2, M3, M4 and M5 subtypes in megacaryocytic differentiated K562 cells whole lysates prepared from K562 cells were analyzed with Western blot technique. Megakaryocytic differentiated K562 cells are homogenized, centrifuged and preserved at -80°C.The protein content of the homogenate are analyzed via Lowry method.Statistically significant differences were determined by using the One-WayAnnova and Bonferroni post test. Here, we show that all muscarinic acetylcholine receptors (M1-M5) are expressed in megakaryocytic differentiated K562 cells. Treatment of megakaryocytic differentiated K562 cells to the cholinergic agonist, CCh, caused an increase in protein expression levels of protein expression pattern M1 and M4 mAChRs in megakaryocytic differentiated K562 cells, which was inhibited by atropine (Fig 1 a,b). Exposure of megakaryocytic differentiated K562 cells to the antimuscarinic agent, atropine, caused increase M2,M3 expression level in 24 h. Conclusions: Five muscarinic receptors were detected on megakaryocytic differentiated K562 cells. Exposure of carbachol increased M1-M4 muscarinic receptor expression. Muscarinic receptors may have a role in megakaryocytic differentiated K562 cells.

Where applicable, experiments conform with Society ethical requirements