Cholinesterases modulate cell adhesion in human neuroblastoma cells in vitro.
Cholinesterases are expressed non-synaptically during embryonic development, neoplasia and neurodegeneration. We have investigated the effects of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and, conversely, anti-AChE and -BChE antibodies and inhibitors on cell adhesion and neurite outgrowth in human neuroblastoma cells. Analysis of cholinesterase levels and isoforms in undifferentiated and differentiated cells indicated a significant rise in AChE levels on differentiation. This increase was related to both cell-associated and secreted enzyme, and was predominantly the G4 isoform. BChE levels and isoforms, on the other hand, showed no significant variation. Coating the tissue culture plate with AChE stimulated neurite outgrowth, while BChE had an anti-adhesive effect. Cell adhesion was affected by the BChE inhibitor, ethopropazine, and the AChE peripheral site inhibitor, BW284c51, but not by eserine which binds to the active site. This indicates that the adhesion function is non-cholinergic, a finding supported by the lack of effect of AE-2, a monoclonal antibody that inhibits AChE, on cell adhesion. Four out of a panel of nine anti-AChE antibodies inhibited adhesion to varying degrees. Of these antibodies, two are catalytic, with epitopes associated with the peripheral anionic site of AChE, and the remaining two have epitopes overlapping this site. Neither of the two anti-BChE antibodies used had any effect on adhesion. These results indicate the importance of AChE in neuroblastoma cell adhesion and neurite outgrowth, and suggest that the peripheral anionic site may be involved in these processes.
Johnson
G
G
Department of Paediatric Surgery and Medical Biochemistry, Faculty of Medicine, University of Stellenbosch, Tygerberg, South Africa. gjo@gerga.sun.ac.za
Moore
S W
SW
D000110:Acetylcholinesterase / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D018680:Cholinergic Antagonists / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology* / Q000201:enzymology*; D009500:Neutralization Tests; D010640:Phenothiazines / Q000494:pharmacology; D011817:Rabbits; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D018680:Cholinergic Antagonists; D010640:Phenothiazines; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; C084820:profenamine; D000110:Acetylcholinesterase