title: Pro-apoptotic properties of morphine in neuroblastoma × glioma NG108-15 hybrid cells: modulation by yohimbine.
abstract: Short-term incubation with pharmacologically relevant concentrations of morphine has been shown to transiently affect the metabolism and redox status of NG108-15 cells through δ-opioid receptor stimulation, but apparently did not provoke cell death. The present work tries to determine if incubation with morphine at longer time intervals (24 h) provokes apoptosis and/or necrosis, as it has been described in other cell lines. We have also checked the potential modulatory role of yohimbine on these effects, on the basis of the previously described interactions between this drug and opioid receptor ligands. Incubation with morphine 0.1 and 10 μM provoked the appearance of images compatible with apoptosis (bebbling, pyknotic cells with cytoplasmic and nuclear condensation) and necrosis (cells swollen with vacuolated cytoplasm lacking cell processes) that could be observed directly and/or after staining with methylene blue, crystal violet and propidium iodide/4',6-diamidino-2-phenylindole (IP/DAPI). Quantification of apoptosis by activation of caspases 3 and 7 and DNA fragmentation with the Tunel assay revealed a modest but significant increase after incubation with the two concentrations of morphine used. Co-incubation with 10 μM yohimbine prevented all these effects of the opioid. The results extend previous findings of a yohimbine-sensitive, neurotoxic effect of morphine on NG108-15 cells.
authors.affiliation: Laboratory of Pharmacology and Toxicology, Universidad San Pablo CEU. Campus Montepríncipe, 28668, Boadilla, Madrid, Spain.
authors.forename: María José
authors.initials: MJ
authors.lastname: Polanco
authors.forename: Luis Fernando
authors.initials: LF
authors.lastname: Alguacil
authors.forename: Carmen
authors.initials: C
authors.lastname: González-Martín
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045744:Cell Line, Tumor; D053938:DNA Fragmentation / Q000187:drug effects; D005910:Glioma / Q000188:drug therapy*; D006822:Hybrid Cells / Q000187:drug effects* / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D051379:Mice; D009020:Morphine / Q000494:pharmacology*; D009336:Necrosis / Q000188:drug therapy / Q000473:pathology; D009447:Neuroblastoma / Q000188:drug therapy*; D017465:Receptors, Opioid, delta / Q000037:antagonists & inhibitors / Q000378:metabolism; D015016:Yohimbine / Q000494:pharmacology*
chemical_list: D017465:Receptors, Opioid, delta; D015016:Yohimbine; D009020:Morphine; C505409:Casp3 protein, mouse; C505524:Casp7 protein, mouse; D053148:Caspase 3; D053179:Caspase 7