Investigation the Effect of Adenosine A1 Receptor Agonist and Antagonist on P53 Gene Expression, and Apoptosis Pathways and Rate in U87Mg Multiform Glioblastoma

Abstract

Background: Improper prognosis in brain cancers requires new treatments. Using family of purinergic receptors with confirmed apoptotic effect can be beneficial. As the role of type A1 receptor in multiform glioblastoma in relation to the P53 gene and apoptotic pathways is nor reported, we studied the role of agonist (N6-cyclopentyladenosine or CPA) and antagonist (8-cyclopentyl-1,3-dipropylxanthine or DPCPX) of this receptor on cell apoptosis and also expression of P53 genes and caspases 7, 8, and 9. Methods: In this study, MTT assay was used to investigate the rate of cellular proliferation, and flowcytometry method with annexin and Pi was also used to investigate early and late cell apoptosis. To evaluate the internal and external apoptotic pathways expression of P53 genes and caspases 7, 8, and 9, real-time reverse transcription polymerase chain reaction (real-time RT PCR) was used. Findings: The treatment of U87Mg cells with DPCPX increased the expression of P53 gene. Expression of caspase 7 as an executive caspase and caspase 9 as a caspase of the mitochondrial pathway of apoptosis increased, but no expression change was observed in the caspase 8 gene. Conclusion: The results of MTT and flowcytometry showed that DPCPX, in addition to suppressing cell proliferation, stimulated apoptosis in U87Mg cells. Inhibition of adenosine A1 receptors by stimulating the expression of genes involved in apoptotic pathways, especially mitochondrial pathway genes, suppressed cell proliferation and induced apoptosis in U87Mg cells.

Keywords

• receptor, adenosine a1
• apoptosis
• p53 genes