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Phosphodiesterase inhibitors control A172 human glioblastoma cell death through cAMP-mediated activation of protein kinase A and Epac1/Rap1 pathways.
Life Sci. 2012 Feb 27; 90(9-10):373-80.LS

Abstract

AIMS

We investigated whether cAMP-mediated protein kinase A(PKA) and Epac1/Rap1 pathways differentially affect brain tumor cell death using 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone(rolipram), specific phosphodiesterase type IV(PDE IV) inhibitor.

MAIN METHODS

A172 and U87MG human glioblastoma cells were used. Percentage of cell survival was determined by MTT assay. PKA and Epac1/Rap1 activation was determined by western blotting and pull-down assay, respectively. Cell cycle and hypodiploid cell formation were assessed by flow cytometry analysis.

KEY FINDINGS

Non-specific PDE inhibitors, isobutylmethylxanthine(IBMX) and theophylline reduce survival percentage of A172 and U87MG cells. The expression of PDE4A and PDE4B was detected in A172 and U87MG cells. Rolipram-treated A172 or U87MG cell survival was lower in the presence of forskolin, adenylate cyclase activator, than that in its absence. Co-treatment with rolipram and forskolin also enhanced CREB phosphorylation on serine 133 that was inhibited by H-89, PKA inhibitor and cAMP-responsive guanine nucleotide exchange factor 1(Epac1), a Rap GDP exchange factor-mediated Rap1 activity in A172 cells. When A172 cells were treated with cell-permeable dibutyryl-cAMP(dbcAMP), PKA activator or 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate(CPT), Epac1 activator, basal level of cell death was increased and cell cycle was arrested at the phase of G2/M. Rolipram-induced A172 cell death was also increased by the co-treatment with dbcAMP or CPT, but it was inhibited by the pre-treatment with H-89.

SIGNIFICANCE

These findings demonstrate that PKA and Epac1/Rap1 pathways could cooperatively play a role in rolipram-induced brain tumor cell death. It suggests that rolipram might regulate glioblastoma cell density through dual pathways of PKA- and Epac1/Rap1-mediated cell death and cell cycle arrest.

Authors+Show Affiliations

Department of Bioscience and Biotechnology, Sejong University, Seoul, Republic of Korea. eunyimoon@sejong.ac.krNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22227470

Citation

Moon, Eun-Yi, et al. "Phosphodiesterase Inhibitors Control A172 Human Glioblastoma Cell Death Through cAMP-mediated Activation of Protein Kinase a and Epac1/Rap1 Pathways." Life Sciences, vol. 90, no. 9-10, 2012, pp. 373-80.
Moon EY, Lee GH, Lee MS, et al. Phosphodiesterase inhibitors control A172 human glioblastoma cell death through cAMP-mediated activation of protein kinase A and Epac1/Rap1 pathways. Life Sci. 2012;90(9-10):373-80.
Moon, E. Y., Lee, G. H., Lee, M. S., Kim, H. M., & Lee, J. W. (2012). Phosphodiesterase inhibitors control A172 human glioblastoma cell death through cAMP-mediated activation of protein kinase A and Epac1/Rap1 pathways. Life Sciences, 90(9-10), 373-80. https://doi.org/10.1016/j.lfs.2011.12.010
Moon EY, et al. Phosphodiesterase Inhibitors Control A172 Human Glioblastoma Cell Death Through cAMP-mediated Activation of Protein Kinase a and Epac1/Rap1 Pathways. Life Sci. 2012 Feb 27;90(9-10):373-80. PubMed PMID: 22227470.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Phosphodiesterase inhibitors control A172 human glioblastoma cell death through cAMP-mediated activation of protein kinase A and Epac1/Rap1 pathways. AU - Moon,Eun-Yi, AU - Lee,Geun-Hee, AU - Lee,Myung-Shik, AU - Kim,Hwan-Mook, AU - Lee,Jae-Wook, Y1 - 2011/12/28/ PY - 2011/09/06/received PY - 2011/11/22/revised PY - 2011/12/13/accepted PY - 2012/1/10/entrez PY - 2012/1/10/pubmed PY - 2012/4/11/medline SP - 373 EP - 80 JF - Life sciences JO - Life Sci VL - 90 IS - 9-10 N2 - AIMS: We investigated whether cAMP-mediated protein kinase A(PKA) and Epac1/Rap1 pathways differentially affect brain tumor cell death using 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone(rolipram), specific phosphodiesterase type IV(PDE IV) inhibitor. MAIN METHODS: A172 and U87MG human glioblastoma cells were used. Percentage of cell survival was determined by MTT assay. PKA and Epac1/Rap1 activation was determined by western blotting and pull-down assay, respectively. Cell cycle and hypodiploid cell formation were assessed by flow cytometry analysis. KEY FINDINGS: Non-specific PDE inhibitors, isobutylmethylxanthine(IBMX) and theophylline reduce survival percentage of A172 and U87MG cells. The expression of PDE4A and PDE4B was detected in A172 and U87MG cells. Rolipram-treated A172 or U87MG cell survival was lower in the presence of forskolin, adenylate cyclase activator, than that in its absence. Co-treatment with rolipram and forskolin also enhanced CREB phosphorylation on serine 133 that was inhibited by H-89, PKA inhibitor and cAMP-responsive guanine nucleotide exchange factor 1(Epac1), a Rap GDP exchange factor-mediated Rap1 activity in A172 cells. When A172 cells were treated with cell-permeable dibutyryl-cAMP(dbcAMP), PKA activator or 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate(CPT), Epac1 activator, basal level of cell death was increased and cell cycle was arrested at the phase of G2/M. Rolipram-induced A172 cell death was also increased by the co-treatment with dbcAMP or CPT, but it was inhibited by the pre-treatment with H-89. SIGNIFICANCE: These findings demonstrate that PKA and Epac1/Rap1 pathways could cooperatively play a role in rolipram-induced brain tumor cell death. It suggests that rolipram might regulate glioblastoma cell density through dual pathways of PKA- and Epac1/Rap1-mediated cell death and cell cycle arrest. SN - 1879-0631 UR - https://www.unboundmedicine.com/medline/citation/22227470/Phosphodiesterase_inhibitors_control_A172_human_glioblastoma_cell_death_through_cAMP_mediated_activation_of_protein_kinase_A_and_Epac1/Rap1_pathways_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0024-3205(11)00601-1 DB - PRIME DP - Unbound Medicine ER -