Cholinergic activation enhances retinoic acid-induced differentiation in the human NB-4 acute promyelocytic leukemia cell line.
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 72h 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.
Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. Electronic address: sadudee.chotirat@gmail.com.
Sadudee
S
Chotirat
Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok 10210, Thailand. Electronic address: tawit@cri.or.th.
Tawit
T
Suriyo
Department of Biomedicine, Faculty of Health, Aarhus University, Aarhus 8000, Denmark. Electronic address: mhokland@biomed.au.dk.
Marianne
M
Hokland
Department of Haematology, Aarhus University Hospital, Aarhus 8000, Denmark. Electronic address: phokland@clin.au.dk.
Peter
P
Hokland
Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok 10210, Thailand; Center of Excellence on Environmental Health and Toxicology, Office of Higher Education Commission, Ministry of Education, Bangkok 10400, Thailand. Electronic address: jutamaad@cri.or.th.
Jutamaad
J
Satayavivad
Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. Electronic address: chirayuaue@yahoo.com.
Chirayu U
CU
Auewarakul
D000109:Acetylcholine / Q000494:pharmacology*; D002217:Carbachol / Q000494:pharmacology; D053148:Caspase 3 / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D018678:Cholinergic Agents / Q000494:pharmacology; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy / Q000473:pathology*; D043587:Receptor, Muscarinic M3 / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects; D014212:Tretinoin / Q000494:pharmacology*
D018678:Cholinergic Agents; D043587:Receptor, Muscarinic M3; D014212:Tretinoin; D002217:Carbachol; C505407:CASP3 protein, human; D053148:Caspase 3; D000109:Acetylcholine
CD11b; Carbachol; Differentiation; NB-4 cells; Non-neuronal cholinergic system