Activation of Cytosolic Phospholipase A2α by Epidermal Growth Factor (EGF) and Phorbol Ester in HeLa Cells: Different Effects of Inhibitors for EGF Receptor, Protein Kinase C, Src, and C-Raf

Mori, Yasuo; Kiuchi, Yoshiaki; Toyomura, Kaori; Matsumoto, Ikiru; Nakamura, Hiroyuki; Fujino, Hiromichi; Murayama, Toshihiko; Kawashima, Tatsuo

doi:10.1254/jphs.09201fp

Cited by 14 publications

(8 citation statements)

References 54 publications

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“…41 Blockage of both results in apoptosis and tumor shrinkage. 42,43 AKT, by inhibiting GSK-3, changes canonical Wnt pathway, releasing ␤-catenin, and leading to angiogenesis and proliferation.…”

Section: Acinar Patternmentioning

confidence: 99%

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Bronchial-pulmonary adenocarcinoma subtyping relates with different molecular pathways

Sousa

Bastos

Silva

et al. 2015

Revista Portuguesa de Pneumologia (English Edition)

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Lung cancer is one of the most common cancers in the world with a high mortality rate. We analyzed 45 surgical samples of the adenocarcinoma, 13 with lymph node metastasis. APC, BCL2, chromogranin A, CK 5/6/18 (LP34), CK20, CK7, cyclin D1, EGFR, ERCC1, HER2, Ki67, LRP, MRP, P53, RB and TTF1 expressions were evaluated by immunohistochemistry (IHC).Higher Ki67, APC, ERCC1 expressions and lower TTF1 expression were identified in advanced stages (IIA and IIIA) of adenocarcinomas, which reflect a more aggressive, less differentiated, possibly a non-TRU adenocarcinoma.Acinar, micropapillary and BA/lepidic adenocarcinoma patterns were the most similar patterns and papillary was the most different pattern followed by solid pattern, according to expression of these markers. Different adenocarcinoma patterns are engaged with different molecular pathways for carcinogenesis, based on the differences of expression. Acinar, BA/lepidic and micropapillary showed higher TTF1 expression (type TRU), and papillary and solid patterns revealed less TTF1 expression, exhibiting a non-TRU/bronchial phenotype. Solid pattern revealed lower HER2 and higher EGFR and ERCC1 (this compared to papillary) expression; papillary higher HER2 and lower ERCC1 expressions; micropapillary higher RB expression; and acinar lower ERCC1 and higher EGFR expressions. Ciclin D1 seems to have more importance in acinar and BA/lepidic patterns than in micropapillary. ERCC1 protein expression in micropapillary, solid and BA/lepidic patterns may indicate DNA repair activation. Inhibition of apoptosis could be explained by BCL2 overexpression, present in all adenocarcinoma patterns. MRP-1 and LRP were overexpressed in all patterns, which may have implications for drug resistance.Further studies are needed to interpret these data regarding to therapy response in advanced staged bronchial-pulmonary carcinomas.

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Section: Acinar Patternmentioning

confidence: 99%

“…Thus, all pathways that this gene regulates are functioning normally or if activated, this activation is not induced by EGFR. 41,57,58 Papillary pattern proliferation properties could in part be explained by APC upregulation when compared to normal tissue.…”

Section: Papillary Patternmentioning

confidence: 99%

Bronchial-pulmonary adenocarcinoma subtyping relates with different molecular pathways

Sousa

Bastos

Silva

et al. 2015

Revista Portuguesa de Pneumologia (English Edition)

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“…The phosphorylation and resulting activation of cPLA 2 α were previously shown to be positively regulated by the MEK‐ERK pathway [Taniguchi et al, ; Tucker et al, ; Leslie et al, ]. U0126 is a selective inhibitor of MEK, an upstream enzyme of ERK, and this reagent at 10 μM almost completely inhibited the MEK/ERK pathway [Taniguchi et al, ; Matsuzawa et al, ]. TNFα‐induced release was significantly decreased in cells pretreated with 10 μM U0126.…”

Section: Resultsmentioning

confidence: 99%

Lactosylceramide‐Induced Phosphorylation Signaling to Group IVA Phospholipase A₂ via Reactive Oxygen Species in Tumor Necrosis Factor‐α‐Treated Cells

Nakamura

Moriyama

Watanabe

et al. 2017

J of Cellular Biochemistry

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The activity of α-type cytosolic phospholipase A (cPLA α, group IVA PLA ), which releases arachidonic acid (AA), is mainly regulated by the Ca -induced intracellular translocation/attachment of the enzyme to substrate membranes and its phosphorylation. We previously reported that tumor necrosis factor-α (TNFα) stimulated the formation of lactosylceramide (LacCer) in L929 fibroblast cells, and this lipid directly bound with and activated cPLA α [Nakamura et al. [2013] J. Biol. Chem. 288:23264-23272]. We herein investigated the role of phosphorylation signaling in the TNFα/LacCer-induced activation of cPLA α in cells. TNFα-treated L929 cells released AA via the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and cPLA α, while a treatment with LacCer alone released AA in a similar manner. The TNFα-induced responses including release of AA were decreased by the inhibition of LacCer synthesis. The treatment with TNFα and LacCer increased the levels of reactive oxygen species (ROS), and the reduction/scavenging of ROS decreased the phosphorylation cascade and release of AA in TNFα/LacCer-treated L929 cells. In the cell line CHO, the treatment with LacCer stimulated the phosphorylation cascade and release of AA via the formation of ROS. Treatments with the anti-LacCer antibody and 4β-phorbol 12-myristate 13-acetate stimulated the phosphorylation cascade, but did not release AA by itself. When combined with the Ca ionophore A23187, treatments with the anti-LacCer antibody and 4β-phorbol 12-myristate 13-acetate released AA. These results, including our previous findings, showed that LacCer alone simultaneously stimulates two processes to activate cPLA α: a phosphorylation signal and attachment of the enzyme to substrate membranes. J. Cell. Biochem. 118: 4370-4382, 2017. © 2017 Wiley Periodicals, Inc.

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“…EGF is known to promote PLA 2 activation [53–55] through several independent signalling pathways [56], that may thus lead to GroPIns4 P [6] and arachidonic acid formation. This is the case for the MEK-dependent phosphorylation cascade that has been involved in both the activation of PLA 2 [57] and formation of GroPIns4 P [6]. Similarly, the activation of PLCγ and cytosolic Ca 2+ increase lead to PLA 2 activation [58].…”

Section: Discussionmentioning

confidence: 99%

A signalling cascade involving receptor-activated phospholipase A2, glycerophosphoinositol 4-phosphate, Shp1 and Src in the activation of cell motility

et al. 2019

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Background Shp1, a tyrosine-phosphatase-1 containing the Src-homology 2 (SH2) domain, is involved in inflammatory and immune reactions, where it regulates diverse signalling pathways, usually by limiting cell responses through dephosphorylation of target molecules. Moreover, Shp1 regulates actin dynamics. One Shp1 target is Src, which controls many cellular functions including actin dynamics. Src has been previously shown to be activated by a signalling cascade initiated by the cytosolic-phospholipase A 2 (cPLA 2 ) metabolite glycerophosphoinositol 4-phosphate (GroPIns4 P ), which enhances actin polymerisation and motility. While the signalling cascade downstream Src has been fully defined, the mechanism by which GroPIns4 P activates Src remains unknown. Methods Affinity chromatography, mass spectrometry and co-immunoprecipitation studies were employed to identify the GroPIns4 P- interactors; among these Shp1 was selected for further analysis. The specific Shp1 residues interacting with GroPIns4 P were revealed by NMR and validated by site-directed mutagenesis and biophysical methods such as circular dichroism, isothermal calorimetry, fluorescence spectroscopy, surface plasmon resonance and computational modelling. Morphological and motility assays were performed in NIH3T3 fibroblasts. Results We find that Shp1 is the direct cellular target of GroPIns4 P . GroPIns4 P directly binds to the Shp1-SH2 domain region (with the crucial residues being Ser 118, Arg 138 and Ser 140) and thereby promotes the association between Shp1 and Src, and the dephosphorylation of the Src-inhibitory phosphotyrosine in position 530, resulting in Src activation. As a consequence, fibroblast cells exposed to GroPIns4 P show significantly enhanced wound healing capability, indicating that GroPIns4 P has a stimulatory role to activate fibroblast migration. GroPIns4 P is produced by cPLA 2 upon stimulation by diverse receptors, including the EGF receptor. Indeed, endogenously-produced GroPIns4 P was shown to mediate the EGF-induced cell motility. Conclusions This study identifies a so-far undescribed mechanism of Shp1/Src modulation that promotes cell motility and that is dependent on the cPLA 2 metabolite GroPIns4 P . We show that GroPIns4 P is required for EGF-induced fibroblast migration and that it is part of a cPLA 2 /GroPIns4 P/ Shp1/Src cascade that might have broad implications for studies of immune-inflammatory response and cancer. El...

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Activation of Cytosolic Phospholipase A2α by Epidermal Growth Factor (EGF) and Phorbol Ester in HeLa Cells: Different Effects of Inhibitors for EGF Receptor, Protein Kinase C, Src, and C-Raf

Cited by 14 publications

References 54 publications

Bronchial-pulmonary adenocarcinoma subtyping relates with different molecular pathways

Bronchial-pulmonary adenocarcinoma subtyping relates with different molecular pathways

Lactosylceramide‐Induced Phosphorylation Signaling to Group IVA Phospholipase A₂ via Reactive Oxygen Species in Tumor Necrosis Factor‐α‐Treated Cells

A signalling cascade involving receptor-activated phospholipase A2, glycerophosphoinositol 4-phosphate, Shp1 and Src in the activation of cell motility

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Activation of Cytosolic Phospholipase A2α by Epidermal Growth Factor (EGF) and Phorbol Ester in HeLa Cells: Different Effects of Inhibitors for EGF Receptor, Protein Kinase C, Src, and C-Raf

Cited by 14 publications

References 54 publications

Bronchial-pulmonary adenocarcinoma subtyping relates with different molecular pathways

Bronchial-pulmonary adenocarcinoma subtyping relates with different molecular pathways

Lactosylceramide‐Induced Phosphorylation Signaling to Group IVA Phospholipase A2 via Reactive Oxygen Species in Tumor Necrosis Factor‐α‐Treated Cells

A signalling cascade involving receptor-activated phospholipase A2, glycerophosphoinositol 4-phosphate, Shp1 and Src in the activation of cell motility

Contact Info

Product

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About

Lactosylceramide‐Induced Phosphorylation Signaling to Group IVA Phospholipase A₂ via Reactive Oxygen Species in Tumor Necrosis Factor‐α‐Treated Cells