Sadudee ChotiratTawit SuriyoMarianne HoklandPeter HoklandJutamaad SatayavivadChirayu U. AuewarakulMahidol UniversityChulabhorn Research InstituteAarhus UniversitetArhus UniversitetshospitalThailand Ministry of Education2018-12-112019-03-142018-12-112019-03-142016-07-01Blood Cells, Molecules, and Diseases. Vol.59, (2016), 77-8410960961107997962-s2.0-84966277160https://repository.li.mahidol.ac.th/handle/20.500.14594/42991© 2016 Elsevier Inc. The non-neuronal cholinergic system (NNCS) has been shown to play a role in regulating hematopoietic differentiation. We determined the expression of cholinergic components in leukemic cell lines by Western blotting and in normal leukocyte subsets by flow cytometry and found a heterogeneous expression of choline acetyltransferase (ChAT), acetylcholinesterase (AChE), choline transporter (CHT), M3 muscarinic acetylcholine receptor (M3-mAChR) and α7 nicotinic acetylcholine receptor (α7-nAChR). We then evaluated NNCS role in differentiation of human NB-4 acute promyelocytic leukemia cell line and discovered a dramatic induction of M3-mAChR after all-trans retinoic acid (ATRA) treatment (p < 0.0001). Adding carbachol which is a cholinergic agonist to the ATRA treatment resulted in an increase of a granulocytic differentiation marker (CD11b) as compared with ATRA treatment alone (p < 0.05), indicating that cholinergic activation enhanced ATRA in inducing NB-4 maturation. The combination of carbachol and ATRA treatment for 72 h also resulted in decreased viability and increased cleaved caspase-3 expression when compared with ATRA treatment alone (p < 0.05). However, this combination did not cause poly (ADP-ribose) polymerase (PARP) cleavage. Overall, we have shown that NB-4 cells expressed M3-mAChR in a differentiation-dependent manner and cholinergic stimulation induced maturation and death of ATRA-induced differentiated NB-4 cells.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1016/j.bcmd.2016.04.009