Induction of apoptosis by an inhibitor of cAMP-specific PDE in malignant murine carcinoma cells overexpressing PDE activity in comparison to their nonmalignant counterparts

Marko, Doris; Romanakis, Konstantinos; Zankl, Heinrich; Fürstenberger, Gerhard; Steinbauer, Brigitte; Eisenbrand, Gerhard

doi:10.1007/bf02737806

Cited by 54 publications

(38 citation statements)

References 27 publications

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“…After synchronization and drug treatment, cells were trypsinized and centrifuged at 200 g. The cell pellet was washed with PBS and fixed in 70% ethanol (4˚C). Flow cytometry was performed as described previously (Marko et al, 1998).…”

Section: Flow Cytometrymentioning

confidence: 99%

Inhibition of cyclin-dependent kinase 1 (CDK1) by indirubin derivatives in human tumour cells

et al. 2001

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SummaryThe bisindole indirubin has been described, more than 30 years ago, as being clinically active in the treatment of human chronic myelocytic leukaemia. However, the underlying mechanism of action has remained unclear. We have reported previously that indirubin and its analogues are potent and selective inhibitors of cyclin-dependent kinases (CDK). In this study, we investigated the influence of indirubin and derivatives on CDK1/cyclin B kinase in human tumour cells at concentrations known to induce growth inhibition. Cells of the mammary carcinoma cell line MCF-7, synchronized by serum deprivation, after serum repletion stay arrested in the G 1 /G 0 phase of the cell cycle in the presence of 2 µM indirubin-3′-monoxime. At higher drug concentrations (≥ 5 µM) an increase of the cell population in the G 2 /M phase is additionally observed. Cells synchronized in G 2 /M phase by nocodazole remain arrested in the G 2 /M phase after release, in the presence of indirubin-3′-monoxime (≥5 µM). After 24 h treatment with 10 µM indirubin-3′-monoxime a sub-G 2 peak appears, indicative for the onset of apoptotic cell death. Treatment of MCF-7 cells with growth inhibitory concentrations of indirubin-3′-monoxime induces dose-dependent inhibition of the CDK1 activity in the cell. After 24 h treatment, a strong decrease of the CDK1 protein level along with a reduction of cyclin B in complex with CDK1 is observed. Taken together, the results of this study strongly suggest that inhibition of CDK activity in human tumour cells is a major mechanism by which indirubin derivatives exert their potent antitumour efficacy.

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Section: Flow Cytometrymentioning

confidence: 99%

Inhibition of cyclin-dependent kinase 1 (CDK1) by indirubin derivatives in human tumour cells

et al. 2001

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“…PDEs are key enzymes in the maintenance of cAMP homeostasis. Many tumour cells have been shown to overexpress cAMP-specific PDEs [16]. Effective PDE inhibition increases the intracellular cAMP level, activating the subsequent protein kinase A (PKA).…”

Section: Introductionmentioning

confidence: 99%

Biological activities of malvidin, a red wine anthocyanidin

Fritz

Kern

Pahlke

et al. 2006

Molecular Nutrition Food Res

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Malvidin (mv) has been identified as a potential inhibitor of 3',5'-cyclic adenosine monophosphate (cAMP) phosphodiesterases (PDE). This study was to investigate if, as a possible consequence of intracellular PDE inhibition, the activity of the mitogen-activated protein kinase (MAPK) cascade is affected by mv treatment. At a concentration of 5 microM of mv a significant decrease of phosphorylated ERK1 and ERK2 (ERK, extracellular regulated kinase) in HT29 cells was observed. However, an increase in substance concentration led to a substantial recurrence of the phosphorylated enzymes. Cell cycle analysis underlined that indeed G(1)-relevant targets are only marginally affected by mv. The recurrence of phosphorylated ERK1/2 and the lack of effectiveness on the G(1)-passage up to 100 microM indicated that the inhibition of cAMP-specific PDEs is of minor relevance for the growth-inhibitory properties of mv in HT29 cells. In contrast, the release of cells, synchronised in the G(2)/M-phase of the cell cycle by nocodazole treatment, was effectively blocked in the presence of 1 microM mv. These results suggest that mv interferes with cellular targets relevant for G(2)/M-progression which have not been identified so far.

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“…[4][5][6][7] The impairment of cyclic adenosine monophosphate (cAMP) or cyclic guanosine monophosphate (cGMP) generation by regulation of phosphodiesterases (PDEs) has been implicated in various cancer pathologies. 8,9 PDEs are enzymes that regulate cellular levels of cAMP/cGMP by controlling their degradation. These enzymes are classified into 11 families (PDE1-PDE11) based on their sequence similarity, substrate preference and sensitivity to various inhibitors.…”

Section: Introductionmentioning

confidence: 99%

Phosphodiesterase-4 promotes proliferation and angiogenesis of lung cancer by crosstalk with HIF

et al. 2012

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Lung cancer is the leading cause of cancer death worldwide. Recent data suggest that cyclic nucleotide phosphodiesterases (PDEs) are relevant in various cancer pathologies. Pathophysiological role of phosphodiesterase 4 (PDE4) with possible therapeutic prospects in lung cancer was investigated. We exposed 10 different lung cancer cell lines (adenocarcinoma, squamous and large cell carcinoma) to hypoxia and assessed expression and activity of PDE4 by real-time PCR, immunocytochemistry, western blotting and PDE activity assays. Expression and activity of distinct PDE4 isoforms (PDE4A and PDE4D) increased in response to hypoxia in eight of the studied cell lines. Furthermore, we analyzed various in silico predicted hypoxia-responsive elements (p-HREs) found in PDE4A and PDE4D genes. Performing mutation analysis of the p-HRE in luciferase reporter constructs, we identified four functional HRE sites in the PDE4A gene and two functional HRE sites in the PDE4D gene that mediated hypoxic induction of the reporter. Silencing of hypoxia-inducible factor subunits (HIF1a and HIF2a) by small interfering RNA reduced hypoxic induction of PDE4A and PDE4D. Vice versa, using a PDE4 inhibitor (PDE4i) as a cyclic adenosine monophosphate (cAMP) -elevating agent, cAMP analogs or protein kinase A (PKA)-modulating drugs and an exchange protein directly activated by cAMP (EPAC) activator, we demonstrated that PDE4-cAMP-PKA/EPAC axis enhanced HIF signaling as measured by HRE reporter gene assay, HIF and HIF target genes expression ((lactate dehydrogenase A), LDHA, (pyruvate dehydrogenase kinase 1) PDK1 and (vascular endothelial growth factor A) VEGFA). Notably, inhibition of PDE4 by PDE4i or silencing of PDE4A and PDE4D reduced human lung tumor cell proliferation and colony formation. On the other hand, overexpression of PDE4A or PDE4D increased human lung cancer proliferation. Moreover, PDE4i treatment reduced hypoxia-induced VEGF secretion in human cells. In vivo, PDE4i inhibited tumor xenograft growth in nude mice by attenuating proliferation and angiogenesis. Our findings suggest that PDE4 is expressed in lung cancer, crosstalks with HIF signaling and promotes lung cancer progression. Thus, PDE4 may represent a therapeutic target for lung cancer therapy.

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Induction of apoptosis by an inhibitor of cAMP-specific PDE in malignant murine carcinoma cells overexpressing PDE activity in comparison to their nonmalignant counterparts

Cited by 54 publications

References 27 publications

Inhibition of cyclin-dependent kinase 1 (CDK1) by indirubin derivatives in human tumour cells

Inhibition of cyclin-dependent kinase 1 (CDK1) by indirubin derivatives in human tumour cells

Biological activities of malvidin, a red wine anthocyanidin

Phosphodiesterase-4 promotes proliferation and angiogenesis of lung cancer by crosstalk with HIF

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