M2 receptor activation inhibits cell cycle progression and survival in human glioblastoma cells.
Bottom Line: Cell growth analysis has demonstrated that the M2 agonist arecaidine strongly decreased cell proliferation in both glioma cell lines and primary cultures.This effect was dose and time dependent.Chemosensitivity assays have, moreover, shown arecaidine and temozolomide similar effects on glioma cell lines, although IC50 value for arecaidine was significantly lower than temozolomide.
Affiliation: Department of Biology and Biotechnologies Charles Darwin, Research Centre of Neurobiology Daniel Bovet, La Sapienza, University of Rome, P.le Aldo Moro, 5-00185 Roma, Italy.Show MeSH
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Mentions: To assess whether or not arecaidine induced apoptotic cell death, we followed two different approaches. First we determined by FACS analysis the fraction of cells with hypodiploid DNA content and higher granulosity (SSC). This fraction was assumed to be apoptotic cells. In Figure 9 we show that arecaidine increased the percentage of U87MG and U251MG hypodiploid cells. U251 cells appeared much more sensitive to the treatment, in fact after 72 hrs of treatment 60% of cells with apoptotic features were found in the U251MG cell lines, whereas in the U87MG cell line, the percentage of apoptotic cells was 14.52% (see Table 4). These data were confirmed by quantifying cytoplasmic nucleosomes, considered to be a hallmark of apoptosis, with an ELISA assay. As shown in Figure 10, treatment with 100 μM arecaidine for 72 hrs induced an increase in the percentage of cytoplasmic nucleosomes both in cell lines (U87 and U251) and in primary cell cultures (CRL-8, MZC-12, FCN-9). The ability of arecaidine to induce apoptosis appears to be higher in glioblastoma cell lines than in primary cultures; in particular U251MG cells displaying the highest sensitivity to apoptosis were arecaidine-induced cells. These data appear in agreement with FACS analysis data.
Affiliation: Department of Biology and Biotechnologies Charles Darwin, Research Centre of Neurobiology Daniel Bovet, La Sapienza, University of Rome, P.le Aldo Moro, 5-00185 Roma, Italy.