GD2 ganglioside-binding antibody 14G2a and specific aurora A kinase inhibitor MK-5108 induce autophagy in IMR-32 neuroblastoma cells
| Autor: | Aneta Wiśniewska, Pawel Pabisz, Iwona Nowak, Katarzyna Wawak, Hanna Rokita, Olga Woźnicka, Małgorzata Durbas, Elżbieta Boratyn, Irena Horwacik |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
autophagy Cancer Research Cyclohexanecarboxylic Acids ganglioside GD2 Clinical Biochemistry Aurora A kinase Pharmaceutical Science Apoptosis Article neuroblastoma Neuroblastoma Mice 03 medical and health sciences Cell Line Tumor Gangliosides Autophagy medicine Animals Humans Cytotoxic T cell Aurora A aurora A Cytotoxicity Aurora Kinase A Cell Proliferation Pharmacology Ganglioside Chemistry Biochemistry (medical) apoptosis Antibodies Monoclonal Cell Biology 14G2a monoclonal antibody medicine.disease In vitro Thiazoles 030104 developmental biology Cancer research Ganglioside GD2 Transcription Factors |
| Zdroj: | Apoptosis |
| ISSN: | 1573-675X 1360-8185 |
| DOI: | 10.1007/s10495-018-1472-9 |
| Popis: | The process of autophagy and its role in survival of human neuroblastoma cell cultures was studied upon addition of an anti-GD2 ganglioside (GD2) 14G2a mouse monoclonal antibody (14G2a mAb) and an aurora A kinase specific inhibitor, MK-5108. It was recently shown that combination of these agents significantly potentiates cytotoxicity against IMR-32 and CHP-134 neuroblastoma cells in vitro, as compared to the inhibitor used alone. In this study we gained mechanistic insights on autophagy in the observed cytotoxic effects exerted by both agents using cytotoxicity assays, RT-qPCR, immunoblotting, and autophagy detection methods. Enhancement of the autophagy process in the 14G2a mAb- and MK-5108-treated IMR-32 cells was documented by assessing autophagic flux. Application of a lysosomotropic agent—chloroquine (CQ) affected the 14G2a mAb- and MK-5108-stimulated autophagic flux. It is our conclusion that the 14G2a mAb (40 μg/ml) and MK-5108 inhibitor (0.1 μM) induce autophagy in IMR-32 cells. Moreover, the combinatorial treatment of IMR-32 cells with the 14G2a mAb and CQ significantly potentiates cytotoxic effect, as compared to CQ used alone. Most importantly, we showed that interfering with autophagy at its early and late step augments the 14G2a mAb-induced apoptosis, therefore we can conclude that inhibition of autophagy is the primary mechanism of the CQ-mediated sensitization to the 14G2a mAb-induced apoptosis. Although, there was no virtual stimulation of autophagy in the 14G2a mAb-treated CHP-134 neuroblastoma cells, we were able to show that PHLDA1 protein positively regulates autophagy and this process exists in a mutually exclusive manner with apoptosis in PHLDA1-silenced CHP-134 cells. Electronic supplementary material The online version of this article (10.1007/s10495-018-1472-9) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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