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: The expression of M2 receptors was investigated in glioblastoma cell cultures and in human fresh glioma specimens. The RT-PCR analysis showed that all stable cell lines (U251MG and U87MG) express the M2 transcript; however, M2 expression in U87MG appeared lower than that in U251MG (Fig. 1A). The real time-PCR analysis showed also that M2 mRNA levels were significantly higher in primary cultures as compared to stable cell lines (Fig. 1B). Western blot analysis confirmed the high expression of M2 receptor in U251MG (Fig. 1C upper panel). Similar results were obtained by immunocytochemistry. Although immunocytochemistry is not a quantitative technique, the intensity of the M2 staining was more evident in U251 (Fig. 2B and D) than in U87 cells (Fig. 2A and C). The amount of M2 receptors revealed in primary cell cultures by Western blot analysis appears in agreement with real time-PCR data (Fig. 1B lower panel).
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.