M2 Muscarinic Receptor Activation Impairs Mitotic Progression and Bipolar Mitotic Spindle Formation in Human Glioblastoma Cell Lines.

Autor: Di Bari M; Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, 00185 Rome, Italy., Tombolillo V; Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, 00185 Rome, Italy., Alessandrini F; Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, 00185 Rome, Italy., Guerriero C; Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, 00185 Rome, Italy., Fiore M; Institute of Molecular Biology and Pathology, CNR, 00185 Rome, Italy., Asteriti IA; Institute of Molecular Biology and Pathology, CNR, 00185 Rome, Italy., Castigli E; Department of Experimental Medicine, Section of Physiology and Biochemistry, University of Perugia, 06100 Perugia, Italy., Sciaccaluga M; Department of Medicine and Surgery, University of Perugia, 06100 Perugia, Italy., Guarguaglini G; Institute of Molecular Biology and Pathology, CNR, 00185 Rome, Italy., Degrassi F; Institute of Molecular Biology and Pathology, CNR, 00185 Rome, Italy., Tata AM; Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, 00185 Rome, Italy.; Research Centre of Neurobiology Daniel Bovet, 00185 Rome, Italy.
Jazyk: angličtina
Zdroj: Cells [Cells] 2021 Jul 08; Vol. 10 (7). Date of Electronic Publication: 2021 Jul 08.
DOI: 10.3390/cells10071727
Abstrakt: Background: Glioblastoma multiforme (GBM) is characterized by several genetic abnormalities, leading to cell cycle deregulation and abnormal mitosis caused by a defective checkpoint. We previously demonstrated that arecaidine propargyl ester (APE), an orthosteric agonist of M2 muscarinic acetylcholine receptors (mAChRs), arrests the cell cycle of glioblastoma (GB) cells, reducing their survival. The aim of this work was to better characterize the molecular mechanisms responsible for this cell cycle arrest.
Methods: The arrest of cell proliferation was evaluated by flow cytometry analysis. Using immunocytochemistry and time-lapse analysis, the percentage of abnormal mitosis and aberrant mitotic spindles were assessed in both cell lines. Western blot analysis was used to evaluate the modulation of Sirtuin2 and acetylated tubulin-factors involved in the control of cell cycle progression.
Results: APE treatment caused arrest in the M phase, as indicated by the increase in p-HH3 (ser10)-positive cells. By immunocytochemistry, we found a significant increase in abnormal mitoses and multipolar mitotic spindle formation after APE treatment. Time-lapse analysis confirmed that the APE-treated GB cells were unable to correctly complete the mitosis. The modulated expression of SIRT2 and acetylated tubulin in APE-treated cells provides new insights into the mechanisms of altered mitotic progression in both GB cell lines.
Conclusions: Our data show that the M2 agonist increases aberrant mitosis in GB cell lines. These results strengthen the idea of considering M2 acetylcholine receptors a novel promising therapeutic target for the glioblastoma treatment.
Databáze: MEDLINE
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