TY  - JOUR
T1  - Effects of atropine on megakaryocytic differentiated K562 leukemia cells
AU  - Cabadak, Hulya
AU  - Kanlı, Zehra
AU  - Aydın, Banu
PY  - 2025
DA  - October
Y2  - 2025
DO  - 10.5472/marumj.1800332
JF  - Marmara Medical Journal
JO  - Marmara Med J
PB  - Marmara University
WT  - DergiPark
SN  - 1019-1941
SP  - 265
EP  - 272
VL  - 38
IS  - 3
LA  - en
AB  - Objective: Non-neuronal cholinergic system signaling pathways play a significant role in various malignancies, including leukemia,lung, colon, brain, and breast cancer. This study aims to investigate the effect of atropine on megakaryocytic differentiated cells, aswell as to identify the apoptosis mechanisms. We also studied the effect of cholinergic drugs on muscarinic receptors and caspase geneexpressions, cell proliferation and caspase activities in differentiated cells.Materials and Methods: K562 cells were induced into megakaryocytic differentiation using phorbol 12-myristate 13-acetate (PMA).The effects of agonists/antagonists on differentiated K562 cells were examined using cell viability and 5-Bromo-2-deoxy-uridine(BrdU) assays. Caspase activities were detected by the caspase assay kit. Protein expression levels were detected by western blotting.Results: Atropine reversed the effects of carbachol (CCh) on increased megakaryocytic differentiated leukemia cell survival andproliferation. The protein expression of M1 and M4 muscarinic receptors was upregulated by CCh, an effect that was reversed byatropine. CCh alone did not significantly change levels of M2, M3, and M5 muscarinic receptor proteins in megakaryocytic differentiatedK562 leukemia cells.Conclusion: M2, M3 muscarinic receptors and caspase 9 may have important functions in preventing the progression of leukemia andmay also make important contributions to targeted therapies in leukemia.
KW  - Atropine
KW  - Leukemia
KW  - Megakaryocytic differentiation
KW  - Muscarinic receptors
KW  - Caspases
CR  - Kawashima K, Fujii T. Extraneuronal cholinergic system in
lymphocytes, Pharmacol Ther 2000;86: 29-48. doi: 10.1016/
s0163-7258(99)00071-6.
CR  - Tobin A B, Budd D C. The anti-apoptotic response of the
Gq/11-coupled muscarinic receptor family. Biochem Soc
Trans 2003; 31: 1182-5. doi: 10.1042/bst0311182.
CR  - Wessler I, Kirkpatrick C J, Racke K. The cholinergic ‘pitfall’:
acetylcholine, a universal cell molecule in biological systems,
including humans. Clin Exp Pharmacol Physiol 1999; 26: 198-
205. doi: 10.1046/j.1440-1681.1999.03016.x
CR  - Lev-Lehman E, Deutsch V, Eldor A, Soreq H. Immature
human megakaryocytes produce nuclear-associated
acetylcholinesterase. Blood 1997;89:3644-53.
CR  - Costa P, Traver D J, Auger C B, Costa L G. Expression of
cholinergic muscarinic receptor subtypes mRNA in rat blood
mononuclear cells. Immunopharmacology 1994;28:113-23.
doi: 10.1016/0162-3109(94)90027-2
CR  - Fukamauchi F, Saunders P A, Hough C, Chuang D M.
Agonist-induced down-regulation and antagonist-induced
up-regulation of m2 – and m3-muscarinic acetylcholine
receptor mRNA and protein in cultured cerebellar granule
cells. Mol Pharmacol 1993;44: 940-9. doi: 10.1111/j.1471-
4159.1991.tb08210.x
CR  - Kawashima K, Fujii T, Moriwaki Y, Misawa H, Horiguchi
K. Reconciling neuronally and nonneuronally derived
acetylcholine in the regulation of immune function.
Ann N Y Acad Sci 2012; 1261: 7-17. doi: 10.1111/j.1749-
6632.2012.06516.x
CR  - Sato K Z, Fujii T, Watanabe Y, et al. Diversity of mRNA
expression for muscarinic acetylcholine receptor subtypes and
neuronal nicotinic acetylcholine receptor subunits in human
mononuclear leukocytes and leukemic cell lines. Neurosci Lett
1999;266:17-20. doi: 10.1016/s0304-3940(99)00259-1
CR  - Paleari L, Grozio A, Cesario A, Russo P. The cholinergic
system and cancer. Semin Cancer Biol 2008;18: 211-7. doi:
10.1016/j.semcancer.2007.12.009
CR  - Spindel E R. Muscarinic receptor agonists and antagonists:
effects on cancer. Handb Exp Pharmacol 2012;208:451-68.
doi: 10.1007/978-3-642-23274-9_19
CR  - Campoy J F, Vidal C J, Munoz-Delgado E, Montenegro M F,
Cabezas-Herrera J, S. Nieto-Ceron S. Cholinergic system and
cell proliferation. Chem Biol Interact 2016;259:257-65. doi:
10.1016/j.cbi.2016.04.014.
CR  - Wessler I, Kirkpatrick C J. Cholinergic signaling controls
immune functions and promotes homeostasis, Int
Immunopharmacol. 2020;83: 106345. doi: 10.1016/j.
intimp.2020.106345
CR  - Cabadak H, Aydin B, Kan B. Regulation of M2, M3, and M4
muscarinic receptor expression in K562 chronic myelogenous
leukemic cells by carbachol. J Recept Signal Transduct Res
2011;31 26-32. doi: 10.3109/10799.893.2010.506484
CR  - Onder Narin G, Aydin B, Cabadak H. Studies on the role
of alpha 7 nicotinic acetylcholine receptors in K562 cell
proliferation and signaling. Mol Biol Rep 2021;48:5045-55.
doi: 10.1007/s11033.021.06498-4
CR  - Jan R, Chaudhry G E. Understanding apoptosis and apoptotic
pathways targeted cancer therapeutics. Adv Pharm Bull
2019;9: 205-18. doi: 10.15171/apb.2019.024
CR  - Boice A, Bouchier-Hayes L. Targeting apoptotic caspases in
cancer. Biochim Biophys Acta Mol Cell Res 2020;1867:118688.
doi: 10.1016/j.bbamcr.2020.118688
CR  - Costa L G, Guizzetti M, Oberdoerster J, et al. Modulation of
DNA synthesis by muscarinic cholinergic receptors. Growth
Factors 2001;18: 227-36. doi: 10.3109/089.771.90109029112
CR  - Lanzafame AA, Christopoulos A, Mitchelson F. Cellular
signaling mechanisms for muscarinic acetylcholine receptors.
Recept Channels 2003;9:241-60.
CR  - Nicke B, Detjen K, Logsdon C D. Muscarinic cholinergic
receptors activate both inhibitory and stimulatory growth
mechanisms in NIH3T3 cells. J Biol Chem 1999;274:21701-6.
doi: 10.1074/jbc.274.31.21701
CR  - Goldman J M, Melo J V. Chronic myeloid leukemia—advances
in biology and new approaches to treatment. N Engl J Med
2003;349: 1451-64. doi: 10.1056/NEJMra020777
CR  - Zhu H Q, Gao F H. Regulatory molecules and corresponding
processes of BCR-ABL protein degradation. J Cancer 2019;10:
2488-500. doi: 10.7150/jca.29528
CR  - Butler T M, Ziemiecki A, Friis R R, Megakaryocytic
differentiation of K562 cells is associated with changes
in the cytoskeletal organization and the pattern of
chromatographically distinct forms of phosphotyrosylspecific
protein phosphatases. Cancer Res 1990;50:6323-
9.
CR  - Aydin B, Cabadak H, Goren M Z. Investigation of the roles of
non-neuronal acetylcholine in chronic myeloid leukemic cells
and their erythroid or megakaryocytic differentiated lines.
Anticancer Agents Med Chem. 2018;18:1440-7. doi: 10.2174
/187.152.0618666.180.406123154
CR  - Lowry O H, Rosebrough N J, Farr A L, Randall R J. Protein
measurement with the Folin phenol reagent. J Biol Chem
1951;193:265-75.
CR  - Lozzio C B, Lozzio B B. Human chronic myelogenous
leukemia cell-line with positive Philadelphia chromosome.
Blood 1975;45:321-34.
CR  - Cabadak H, Kucukibrahimoglu E, Aydin B, Kan B, Zafer
Goren M. Muscarinic receptor-mediated nitric oxide release in
a K562 erythroleukaemia cell line. Auton Autacoid Pharmacol
2009;29: 109-15. doi: 10.1111/j.1474-8673.2009.00431.x
CR  - Jabbour E, Cortes J E, Ghanem H, O’Brien S, Kantarjian
H M. Targeted therapy in chronic myeloid leukemia.
Expert Rev Anticancer Ther 2008;8: 99-110. doi:
10.1586/14737140.8.1.99
CR  - Song P, Sekhon H S, Lu A, et al. M3 muscarinic receptor
antagonists inhibit small cell lung carcinoma growth and
mitogen-activated protein kinase phosphorylation induced
by acetylcholine secretion. Cancer Res 2007;67: 3936-44. doi:
10.1158/0008-5472.CAN-06-2484
CR  - Ami A, Koga K, Fushiki H, Ueno Y, OginoY, Ohta H. Selective
M3 muscarinic receptor antagonist inhibits small-cell lung
carcinoma growth in a mouse orthotopic xenograft model. J
Pharmacol Sci. 2011;116: 81-8. doi: 10.1254/jphs.10308fp
CR  - Rosemond E, Rossi M, McMillin S M, Scarselli M, Donaldson
J G, Wess J. Regulation of M(3) muscarinic receptor expression
and function by transmembrane protein 147. Mol Pharmacol
2011;79: 251-61. doi: 10.1124/mol.110.067363
CR  - Degirolamo C, Modica S, Palasciano G, Moschetta A. Bile
acids and colon cancer: Solving the puzzle with nuclear
receptors. Trends Mol Med 2011;17:564-72. doi: 10.1016/j.
molmed.2011.05.010
CR  - Gutkind J S, Novotny E A, Brann M R, Robbins K C.
Muscarinic acetylcholine-receptor subtypes as agonistdependent
oncogenes, P Natl Acad Sci 1991; 88: 4703-7. doi:
10.1073/pnas.88.11.4703
CR  - Baumgold J, Dyer K. Muscarinic receptor-mediated inhibition
of mitogenesis via a protein-kinase C-independent mechanism
in M1-T-transfected A9 L-cells, Cell Signal 1994;6:103-8. doi:
10.1016/0898-6568(94)90065-5
CR  - Renz B W, Tanaka T, Sunagawa M, et al. Cholinergic signaling
via muscarinic receptors directly and indirectly suppresses
pancreatic tumorigenesis and cancer stemness. Cancer Discov
2018;8: 1458-73. doi: 10.1158/2159-8290.CD-18-0046
CR  - Bennett JA , Ture S K, Schmidt RA, et al. Acetylcholine inhibits
platelet activation. Pharmacol Exp Ther 2019;369:182-7. doi:
10.1124/jpet.118.253583
CR  - Reina S, Sterin-Borda L, Passafaro D, Borda E. Muscarinic
cholinoceptor activation by pilocarpine triggers apoptosis in
human skin fibroblast cells. J Cell Physiol 2010;222:640-7. doi:
10.1002/jcp.21981
CR  - Ahmed E A, Alkuwayti M A, Ibrahim H M. Atropine is a
suppressor of epithelial-mesenchymal transition (EMT) that
reduces stemness in drug-resistant breast cancer cells. Int J
Mol Sci 2022;23:9849. doi: 10.3390/ijms23179849
UR  - https://doi.org/10.5472/marumj.1800332
L1  - https://dergipark.org.tr/en/download/article-file/5315891
ER  -