PCPH/ENTPD5 Expression Enhances the Invasiveness of Human Prostate Cancer Cells by a Protein Kinase Cδ–Dependent Mechanism

Villar, Joaquín; Arenas, María Isabel; MacCarthy, Caitlin M.; Blánquez, M. J.; Tirado, Óscar M.; Notario, Vicente

doi:10.1158/0008-5472.can-07-2041

Cited by 39 publications

(56 citation statements)

References 44 publications

(64 reference statements)

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“…There is also evidence that PKCd can enhance cell motility, possibly by affecting the expression and activity of b1 integrins and focal adhesion kinase (Chen et al 2007, Brenner et al 2008. Furthermore, PKCd has been shown to act to promote invasiveness in prostate and breast cancer cells (Kiley et al 1999, Kharait et al 2006, Villar et al 2007, and rottlerin has been shown to completely prevent invasion induced by the integrin-activating peptide PHSRN in DU-125 prostate cancer cells (Zeng et al 2006). We have shown that both FSH and LH induce migration in ovarian cancer cells that is dependent upon ERK1/2 and PKCd signalling.…”

Section: Discussionmentioning

confidence: 99%

Gonadotropin-induced ovarian cancer cell migration and proliferation require extracellular signal-regulated kinase 1/2 activation regulated by calcium and protein kinase Cδ

Mertens‐Walker¹,

Bolitho²,

Baxter³

et al. 2010

Endocrine-Related Cancer

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The gonadotropin hypothesis proposes that elevated serum gonadotropin levels may increase the risk of epithelial ovarian cancer (EOC). We have studied the effect of treating EOC cell lines (OV207 and OVCAR-3) with FSH or LH. Both gonadotropins activated the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase 1/2 (ERK1/2) pathway and increased cell migration that was inhibited by the MAPK 1 inhibitor PD98059. Both extra-and intracellular calcium ion signalling were implicated in gonadotropin-induced ERK1/2 activation as treatment with either the calcium chelator EGTA or an inhibitor of intracellular calcium release, dantrolene, inhibited gonadotropin-induced ERK1/2 activation. Verapamil was also inhibitory, indicating that gonadotropins activate calcium influx via L-type voltage-dependent calcium channels. The cAMP/protein kinase A (PKA) pathway was not involved in the mediation of gonadotropin action in these cells as gonadotropins did not increase intracellular cAMP formation and inhibition of PKA did not affect gonadotropin-induced phosphorylation of ERK1/2. Activation of ERK1/2 was inhibited by the protein kinase C (PKC) inhibitor GF 109203X as well as by the PKCd inhibitor rottlerin, and downregulation of PKCd was inhibited by small interfering RNA (siRNA), highlighting the importance of PKCd in the gonadotropin signalling cascade. Furthermore, in addition to inhibition by PD98059, gonadotropin-induced ovarian cancer cell migration was also inhibited by verapamil, GF 109203X and rottlerin. Similarly, gonadotropin-induced proliferation was inhibited by PD98059, verapamil, GF 109203X and PKCd siRNA. Taken together, these results demonstrate that gonadotropins induce both ovarian cancer cell migration and proliferation by activation of ERK1/2 signalling in a calcium-and PKCd-dependent manner.

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Section: Discussionmentioning

confidence: 99%

Gonadotropin-induced ovarian cancer cell migration and proliferation require extracellular signal-regulated kinase 1/2 activation regulated by calcium and protein kinase Cδ

Mertens‐Walker¹,

Bolitho²,

Baxter³

et al. 2010

Endocrine-Related Cancer

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“…Our studies therefore support a positive role for PKCδ in the malignant progression of tumors. Actually, elevated expression of PKCδ has been detected in certain tumors Nabha et al, 2005;Villar et al, 2007) and has been found to increase the motility and metastatic potential of tumor cells (Kiley et al, 1999; Alonso-Escolano et al, 2006;Kharait et al, 2006;Villar et al, 2007).…”

Section: Journal Of Cell Sciencementioning

confidence: 99%

“…Increasing evidence also indicates that PKCδ has a positive role in cell motility (Chen et al, 2007;Gliki et al, 2002;Iwabu et al, 2004;Li et al, 2003) and the metastatic potential of tumor cells (Kiley et al, 1999;Kruger and Reddy, 2003; Alonso-Escolano et al, 2006;Kharait et al, 2006;Villar et al, 2007). However, the role of PKCδ in intercellular junctions remains obscure.…”

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confidence: 99%

Functional suppression of E-cadherin by protein kinase Cδ

Chen

2009

Journal of Cell Science

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Journal of Cell Science 514 cell functions, including cell proliferation (Ashton et al., 1999;Kitamura et al., 2003), differentiation (Corbit et al., 1999;Pessino et al., 1995), apoptosis (Brodie and Blumberg, 2003;Kajimoto et al., 2004;Zhong et al., 2002) and tumor suppression (Lu et al., 1997;Reddig et al., 1999). Increasing evidence also indicates that PKCδ has a positive role in cell motility (Chen et al., 2007;Gliki et al., 2002;Iwabu et al., 2004;Li et al., 2003) and the metastatic potential of tumor cells (Kiley et al., 1999;Kruger and Reddy, 2003; Alonso-Escolano et al., 2006;Kharait et al., 2006;Villar et al., 2007). However, the role of PKCδ in intercellular junctions remains obscure. In this study, we aim to explore the role of PKCδ in cellcell junctions using Madin-Darby canine kidney (MDCK) cells as a model. Results Expression of GFP-PKCδ, but not GFP-PKCα, suppresses the homophilic interactions between the ectodomains of E-cadherins in MDCK cellsTo explore the role of PKC in cell-cell junctions, GFP-PKCδ and GFP-PKCα were stably expressed in MDCK cells (Fig. 1A). Expression of either construct in those cells did not alter their growth (data not shown) or their ability to form cell colonies within which the cells are in contact with each other (Fig. 1B). To examine their effect on adherens junctions and tight junctions, cells stably expressing GFP-PKCδ or GFP-PKCα were co-cultured with parental control MDCK cells and then grown to confluence. The cells were fixed and stained for adherens junctions and tight junctions with anti-E-cadherin and anti-ZO-1, respectively. Neither GFP-PKCδ nor GFP-PKCα affected tight junctions of MDCK cells (Fig. 1C,D). However, the fluorescence intensity of E-cadherin at cell-cell contacts in the cells expressing GFP-PKCδ, but not GFP-PKCα, was much weaker than that in adjacent parental MDCK cells (Fig. 1C,D).As the total and cell surface levels of E-cadherins were not altered by GFP-PKCδ (supplementary material Fig. S1), it is unlikely that the decreased fluorescence intensity of E-cadherin was due to suppression of E-cadherin expression. The rat monoclonal anti-Ecadherin (clone ECCD-2) used in Fig. 1C is known to recognize the extracellular domain of E-cadherin, rendering it possible that GFP-PKCδ causes a conformational change on the ectodomain of E-cadherin, which then prevents E-cadherins from detection by the ECCD-2 antibody. To clarify this, a mouse monoclonal antibody (clone 36) that recognizes the cytoplasmic portion of E-cadherins was used. The fluorescence intensity of E-cadherin with the clone 36 antibody did not show differences between control cells and PKCδ-overexpressed cells ( Fig. 2A), supporting the idea that the conformation of E-cadherins rather than their distribution or expression is affected by PKCδ. In addition, we found that the ability of the ECCD-2 antibody, but not the clone 36 antibody, to detect E-cadherin at cell-cell junctions relied on the presence of Ca 2+ in the culture medium, as demonstrated by Ca 2+ -switch assay (Fig. 2B), indicat...

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“…Previous studies have shown that invasion of prostate cancer cells (PC-3) is dependent on the activity of different PKCs, among them PKC␦ (46,49). PC-3 cells expressing Spry2 were shown to be less invasive, comparable with those cells where endogenous PKC␦ was knocked down or where the activity of endogenous PKC␦ was blocked by a kinase-dead, dominant negative PKC␦.…”

Section: Discussionmentioning

confidence: 99%

“…In prostate cancer cells, PKC␦ has been shown to be important for cell invasion (46). To gain insight into the physiological relevance of this interaction, we investigated whether Spry2 could affect the invasiveness of a prostate cancer cell line, PC-3.…”

Section: Spry2 Inhibits Cell Invasion Of Pc-3 Cells-pkc␦ Has Beenmentioning

confidence: 99%

Sprouty2 Interacts with Protein Kinase Cδ and Disrupts Phosphorylation of Protein Kinase D1

Chow

Guy

2009

Journal of Biological Chemistry

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The Sprouty (Spry) proteins act as inhibitors of the Ras/ERK pathway downstream of receptor tyrosine kinases. In this study, we report a novel interaction between protein kinase C ␦ (PKC␦) and Spry2. Endogenous PKC␦ and Spry2 interact in cells upon basic fibroblast growth factor stimulation, indicating a physiological relevance for the interaction. This interaction appeared to require the full-length Spry2 protein and was conformationdependent. Conformational constraints were released upon FGFR1 activation, allowing the interaction to occur. Although this interaction did not affect the phosphorylation of PKC␦ by another kinase, it reduced the phosphorylation of a PKC␦ substrate, protein kinase D1 (PKD1). Spry2 was found to interact more strongly with PKC␦ with increasing amounts of PKD1, which indicated that instead of competing with PKD1 for binding with PKC␦, it was more likely to form a trimeric complex with both PKC␦ and PKD1. Formation of the complex was found to be dependent on an existing PKC␦-PKD1 interaction. By disrupting the interaction between PKC␦ and PKD1, Spry2 was unable to associate with PKC␦ to form the trimeric complex. As a consequence of this trimeric complex, the existing interaction between PKC␦ and PKD1 was increased, and the transfer of phosphate groups from PKC␦ to PKD1 was at least partly blocked by Spry2. The action of Spry2 on PKC␦ resulted in the inhibition of both ERK phosphorylation and invasion of PC-3 cells via PKC␦ signaling. By disrupting the capacity of PKC␦ to phosphorylate its cognate substrates, Spry2 may serve to modulate PKC␦ signaling downstream of receptor tyrosine kinases and to regulate the physiological outcome.

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PCPH/ENTPD5 Expression Enhances the Invasiveness of Human Prostate Cancer Cells by a Protein Kinase Cδ–Dependent Mechanism

Cited by 39 publications

References 44 publications

Gonadotropin-induced ovarian cancer cell migration and proliferation require extracellular signal-regulated kinase 1/2 activation regulated by calcium and protein kinase Cδ

Gonadotropin-induced ovarian cancer cell migration and proliferation require extracellular signal-regulated kinase 1/2 activation regulated by calcium and protein kinase Cδ

Functional suppression of E-cadherin by protein kinase Cδ

Sprouty2 Interacts with Protein Kinase Cδ and Disrupts Phosphorylation of Protein Kinase D1

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