olanzapine and Neuroblastoma

Recent clinical studies have suggested that treatment with second generation antipsychotic drugs such as olanzapine may prevent progressive alterations of brain structure in patients with schizophrenia. However, the molecular mechanisms underlying these different effects remain to be determined. We investigated the mechanisms of action of olanzapine and haloperidol, on serum withdrawal apoptosis in human neuroblastoma SH-SY5Y cells.. SH-SY5Y cells were cultured with olanzapine and haloperidol in medium with or without serum. We determined the effects of the drugs on cell viability against serum withdrawal by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Additionally, to explore the drugs' actions, Western blot was performed to examine the expression of key genes involved in GSK-3beta-mediated signaling, notably GSK-3beta, beta-catenin, and Bcl-2.. SH-SY5Y cells suffered about a 38% loss in cell number under serum-free conditions for 48 h. Olanzapine (10-200 muM) up to 100 muM significantly attenuated serum withdrawal-induced cell loss (p<0.01), and a dose of 100 muM also increased cell viability (p<0.05). In contrast, haloperidol (0.01-10 muM) did not affect cell viability but exacerbated cell death at 10 muM under serum-free conditions (p<0.01). Western blot analysis showed that olanzapine, but not haloperidol, prevented the serum withdrawal-induced decrease in levels of neuroprotective proteins such as p-GSK-3beta, beta-catenin, and Bcl-2 (p<0.01), whereas haloperidol robustly reduced the levels of these proteins at a 10 muM dose in serum-starved cells (p<0.05). Moreover, olanzapine alone significantly increased phosphorylation of GSK-3beta under normal conditions (p<0.05).. This study showed that olanzapine may have neuroprotective effects, whereas haloperidol was apparently neurotoxic. The actions of signaling systems associated with GSK-3beta may be key targets for olanzapine and haloperidol, but their effects are distinct. These differences suggest different therapeutic effects of first and second generation antipsychotic drugs in patients with schizophrenia.

Topics: Analysis of Variance; Antipsychotic Agents; Apoptosis; Benzodiazepines; beta Catenin; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Gene Expression Regulation; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Haloperidol; Humans; Neuroblastoma; Olanzapine; Proto-Oncogene Proteins c-bcl-2; Serum; Tetrazolium Salts; Thiazoles

Olanzapine has previously been shown to stimulate the growth of neuronal cells in culture. A major goal of the present studies was to determine if olanzapine also provided neuroprotection to pheochromocytoma (PC12) cells, SH-SY5Y neuroblastoma cells, and primary cultures of rat cortical neurons. Olanzapine was mitogenic and enhanced the survival of PC12 cells, SH-SY5Y cells and 3T3 preadipocytes, but not L6 myoblasts or myeloma cells. It protected neuronal cells from death induced by serum and glutamine deprivation, amyloid beta peptide (25-35), and fluphenazine. Molecular mechanisms of the neuroprotection by olanzapine were explored, specifically the activation of various protein kinase signaling pathways including Akt/protein kinase B (PKB), extracellular-regulated kinase (ERK), ERK1/2, and mitogen-activated protein kinase (MAPK), p38. Olanzapine treatment led to rapid phosphorylation of kinases from all three pathways in PC12 cells. Phosphorylation of Akt was blocked with selective inhibitors (wortmannin and LY294002), which implicates phosphoinositide 3-kinase (PI3K) in the signaling cascade. Short-term mitogenic effects of olanzapine were abolished with a selective inhibitor of Akt, but not by inhibition of the ERK pathway. Other antipsychotic drugs stimulated phosphorylation of a subset of the kinase panel, but not all three kinases. The present findings demonstrate that olanzapine has both mitogenic and neuroprotective effects in neuronal cells.

Topics: 3-Phosphoinositide-Dependent Protein Kinases; Amyloid beta-Peptides; Animals; Antipsychotic Agents; Benzodiazepines; Blotting, Western; Bromodeoxyuridine; Cell Division; Cells, Cultured; Cerebral Cortex; Culture Media, Serum-Free; Dose-Response Relationship, Drug; Drug Interactions; Enzyme Activation; Enzyme Inhibitors; Humans; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Neuroblastoma; Neurons; Olanzapine; p38 Mitogen-Activated Protein Kinases; Peptide Fragments; Pertussis Toxin; Phosphorylation; Protein Serine-Threonine Kinases; Rats; Tetrazolium Salts; Thiazoles; Time Factors

In radioligand binding studies, BIMG 80, a new putative antipsychotic, displayed good affinity at certain serotonin (5-HT1A, 5-HT2A, 5-HT6), dopamine (D1, D2L, D4), and noradrenergic (alpha1) receptors. The effect of acute subcutaneous BIMG 80, clozapine, haloperidol, risperidone, amperozide, olanzapine, and Seroquel was then investigated on dopamine release in medial prefrontal cortex, nucleus accumbens, and striatum in freely moving rats using the microdialysis technique. Four different neurochemical profiles resulted from the studies: (a) Systemic administration of BIMG 80, clozapine, and amperozide produced greater percent increases in dopamine efflux in medial prefrontal cortex than in the striatum or the nucleus accumbens. (b) Haloperidol induced a similar increase in dopamine concentrations in the striatum and nucleus accumbens with no effect in the medial prefrontal cortex. (c) Risperidone and olanzapine stimulated dopamine release to a similar extent in all brain regions investigated. (d) Seroquel failed to change significantly dopamine output both in the medial prefrontal cortex and in the striatum. Because an increase in dopamine release in the medial prefrontal cortex may be predictive of effectiveness in treating negative symptoms and in the striatum may be predictive of induction of extrapyramidal side effects, BIMG 80 appears to be a potential antipsychotic compound active on negative symptoms of schizophrenia with a low incidence of extrapyramidal side effects.

Topics: Animals; Antipsychotic Agents; Benzodiazepines; CHO Cells; Clozapine; Corpus Striatum; COS Cells; Cricetinae; Dibenzothiazepines; Dopamine; Female; Guinea Pigs; Haloperidol; Indoles; Kidney; Male; Microdialysis; Neuroblastoma; Nucleus Accumbens; Olanzapine; Piperazines; Pirenzepine; Prefrontal Cortex; Pyridines; Quetiapine Fumarate; Radioligand Assay; Rats; Rats, Sprague-Dawley; Risperidone; Tritium; Tumor Cells, Cultured