A Ligand-inducible Epidermal Growth Factor Receptor/Anaplastic Lymphoma Kinase Chimera Promotes Mitogenesis and Transforming Properties in 3T3 Cells

Piccinini, Giulia; Bacchiocchi, Roberta; Serresi, Michela; Vivani, C.; Rossetti, Silvia; Gennaretti, Claudia; Carbonari, Damiano; Fazioli, Francesca

doi:10.1074/jbc.m111145200

Cited by 28 publications

(28 citation statements)

References 54 publications

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“…A recent paper using an EGF-activated EGFR-ALK chimera suggests that the MAPK cascade is only very weakly induced in response to an activation of the ALK tyrosine kinase (29). This may be due to the fact that this chimeric receptor is not fully activated in response to EGF.…”

Section: Discussionmentioning

confidence: 99%

Anti-apoptotic Signaling of Pleiotrophin through Its Receptor, Anaplastic Lymphoma Kinase

Bowden

Stoica

Wellstein

2002

Journal of Biological Chemistry

102

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The secreted growth factor pleiotrophin (PTN) can induce mitogenesis in cells that express the receptor for this growth factor, anaplastic lymphoma kinase (ALK). Here we examine the ability of PTN to produce antiapoptotic signals. We demonstrate that PTN is a survival factor for SW-13 epithelial cells and show that ribozymemediated depletion of ALK from SW-13 cells abolishes this effect of PTN. Furthermore, in serum-starved NIH3T3 fibroblasts PTN prevents apoptosis (measured by annexin V staining) with an EC 50 of 0.2 ng/ml and induces cell growth at higher concentrations of PTN. A polyclonal antibody against the PTN ligand-binding domain of the ALK receptor (␣-LBD) was a partial agonist for ALK in NIH3T3 cells. This ␣-LBD antibody showed high agonist activity for anti-apoptosis (56 ؎ 9% relative to PTN), low agonist activity for cell growth (21 ؎ 1% relative to PTN), and was an antagonist of PTN-induced cell growth (61 ؎ 2% inhibition). Both MAP kinase and phosphatidylinositol (PI) 3-kinase cascades in NIH3T3 cells were activated by PTN, and this effect persisted for up to 3 h. Surprisingly, the anti-apoptotic effect of PTN was completely blocked by the MAP kinase inhibitor UO126, but was not affected by the PI 3-kinase inhibitor LY294002. In contrast, PTN-dependent cell growth required both MAPK and PI 3-kinase activity. We conclude that anti-apoptotic signaling of PTN through ALK in NIH3T3 fibroblasts is via the MAP kinase pathway.The ability of multicellular organisms to maintain homeostasis is dependent upon a delicate balance between cell survival and programmed cell death or apoptosis. One of the key elements regulating this balance is growth factor signaling. Growth factors and cytokines have been shown to regulate a wide variety of cellular effects, including cell survival. Growth factor-mediated effects occur through the stimulation of specific intracellular signaling cascades via the intrinsic tyrosine kinase activities of their receptors. For tyrosine kinase receptors, ligand-dependent activation results in autophosphorylation of tyrosine residues within the cytosolic domain of the receptor creating specific binding sites for intracellular signal transduction molecules.

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Section: Discussionmentioning

confidence: 99%

Anti-apoptotic Signaling of Pleiotrophin through Its Receptor, Anaplastic Lymphoma Kinase

Bowden

Stoica

Wellstein

2002

Journal of Biological Chemistry

102

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“…Lysates or immunocomplexes were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred onto nitrocellulose filters, and immunoblotted according to previously described procedures. [26][27] Bound proteins were visualized with…”

Section: Cell Lysis and Protein Analysismentioning

confidence: 99%

“…We have previously shown that in this chimera the biochemical and biologic properties of ALK are controlled by EGF stimulation, thus making it possible to dissect ALK enzymatic function under condition of controlled ligand-induced activation. 26 Therefore, fibroblastic NIH-3T3 cells expressing the chimera EGFR/ALK (NIH-EGFR/ALK) were cultured at 37°C for different time periods in the absence or presence of EGF prior to lysis and subsequent in vitro DGK assay on total homogenate. As shown in Figure 2, an increase in total DGK activity was observed after EGF treatment, further supporting a direct correlation between ALK activation and stimulation of a cellular DGK enzymatic function.…”

Section: Coupling Of Alk With Dg Kinase Activitymentioning

confidence: 99%

“…NPM/ALK-infected 32D cells, the human NPM/ALK-negative T lymphoblastoid CEM cell line, the human NPM/ALK-positive Karpas 299 (kindly provided by B. Falini, University of Perugia, Italy), and TS (kindly provided by R. Piva, University of Turin, Italy) cell lines were cultured in RPMI 1640 containing 10% FBS. Genetically engineered NIH-3T3 cells overexpressing the chimeric epidermal growth factor receptor (EGFR)-ALK receptor (NIH-EGFR/ALK) were described previously 26 and maintained in Dulbecco modified Eagle medium (D-MEM; Invitrogen) containing 10% FBS. For epidermal growth factor (EGF) triggering experiments, subconfluent NIH-EGFR/ALK cell monolayers, grown in poly-L-lysine-coated dishes, were incubated for 18 hours in D-MEM medium supplemented with transferrin (5 g/mL; Becton Dickinson, Franklin Lakes, NJ) and Na 2 SeO 3 (10 Ϫ8 M; Sigma) in the absence of serum (starvation conditions) before EGF treatment (Upstate Biotechnology, Lake Placid, NY).…”

Section: Cell Lines and Transfectionsmentioning

confidence: 99%

“…Total cell lysates were prepared at 4°C by using 1% Triton X-100 lysis buffer as previously described, 26 unless otherwise specified. For immunoprecipitation procedures, cellular lysates were incubated with the indicated antibody for at least 2 hours at 4°C and immunocomplexes were recovered by adsorption to Protein-G Sepharose beads (Amersham Biosciences, Piscataway, NJ).…”

Section: Cell Lysis and Protein Analysismentioning

confidence: 99%

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Activation of -diacylglycerol kinase is critical for the mitogenic properties of anaplastic lymphoma kinase

Bacchiocchi

Baldanzi

Carbonari

et al. 2005

Blood

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IntroductionThe anaplastic lymphoma kinase (ALK) gene, localized on chromosome 2p23, encodes for a large, glycosylated 200-kDa membranespanning tyrosine kinase receptor whose expression is highly tissue specific, being restricted to components of the nervous system; structural homology studies place ALK within the family of insulin receptor tyrosine kinases (RTKs), with the highest homology to the leukocyte tyrosine kinase (LTK). 1,2 Involvement of ALK in the pathogenesis of hematopoietic malignancies derives from the observation that constitutively active forms of ALK are detected with a high frequency in anaplastic large-cell lymphomas (ALCLs), a subgroup of non-Hodgkin lymphomas, predominantly of T or null type. 3 The oncogenic forms of ALK are the result of somatic chromosome translocations that fuse the ALK cytoplasmic domain to the 5Ј region from different partner genes. The most frequent oncogenic version of ALK is represented by nucleophosmin (NPM)/ALK, an 80-kDa hybrid protein created by the t(2;5)(p23; q35) rearrangement. 4,5 The tumorigenic properties of NPM/ALK have been demonstrated in vitro in different cell systems [6][7][8][9] and confirmed in vivo by the generation of NPM/ALK-mediated tumor models. [10][11][12][13] Despite several studies, the pathogenic mechanisms leading to ALK-mediated transformation remain still poorly defined.Thus far, several signaling molecules have been identified that associate and/or are activated by ALK, including growth factor receptor-bound protein 2 (Grb2), Src homology and collagen (Shc), insulin receptor substrate-1 (IRS-1), phospholipase C-␥ (PLC-␥), p60 src , and phosphatidylinositol 3-kinase (PI3-K), although only a few have been shown to be strictly specific for ALK's transforming potential. 3 In particular, several lines of investigation have indicated that both PLC-␥ and PI3-K are critical transducers of NPM/ALK-mediated oncogenesis through activation of mitogenic and/or survival signals, 8,[14][15] while the exact role of the ras/mitogen activated protein kinase (MAPK) cascade needs further evaluation. 3 The contribution of p60 src has been recently evaluated in NPM/ALK-positive cell lines and demonstrated through the effects of p60 src down-regulation and pharmacologic inhibition on cellular proliferative rate. 16 Additional relevant effectors of NPM/ALK-mediated lymphomagenesis are represented by signal transducer and activator of transcription 3 (Stat3) and Stat5. 17 Obviously, further investigations are needed to identify other pathways that are operative in ALK-positive malignancies and that could represent intracellular targets for a more appropriated therapeutic intervention in ALCL treatment. For personal use only. on May 11, 2018. by guest www.bloodjournal.org From Diacylglycerol kinase (DGK) has attracted much attention in recent years since growing evidence indicates its direct involvement in the regulation of signal transduction processes. The family of mammalian DGKs comprises 9 isoenzymes classified in 5 distinct groups on the basis of their ...

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Anaplastic lymphoma kinase is dynamically expressed on subsets of motor neurons and in the peripheral nervous system

et al. 2006

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During embryonic development, complex events such as cellular proliferation, differentiation, survival, and guidance of axons are orchestrated and regulated by a variety of extracellular signals. Receptor tyrosine kinases mediate many of these events with several playing critical roles in neuronal survival and axonal guidance. It is evident that not all the receptor tyrosine kinases that play key roles in regulating neuronal development have been identified. In this study, we have characterized the spatial-temporal expression profile of a recently identified receptor tyrosine kinase, anaplastic lymphoma kinase (ALK), in embryonic chick by means of whole mount in situ hybridization in conjunction with immunohistochemistry. Our findings reveal that Alk is expressed in sympathetic and dorsal root ganglia as early as stage 19. In addition, mRNA is expressed from stage 23/24 (E4) until stage 39 (E13) in discrete motor neuron subsets of chick spinal cord along with a select group of muscles that are innervated by one of these particular motor neuron clusters. Interestingly, expression within the spinal cord is coincident with the onset and duration of motor neuron programmed cell death and during the period of musculature innervation and synapse formation. Hence, the data presented here identify ALK as a novel candidate receptor for regulating critical events in the development of neurons in both the central and peripheral nervous system.

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A Ligand-inducible Epidermal Growth Factor Receptor/Anaplastic Lymphoma Kinase Chimera Promotes Mitogenesis and Transforming Properties in 3T3 Cells

Cited by 28 publications

References 54 publications

Anti-apoptotic Signaling of Pleiotrophin through Its Receptor, Anaplastic Lymphoma Kinase

Anti-apoptotic Signaling of Pleiotrophin through Its Receptor, Anaplastic Lymphoma Kinase

Activation of -diacylglycerol kinase is critical for the mitogenic properties of anaplastic lymphoma kinase

Anaplastic lymphoma kinase is dynamically expressed on subsets of motor neurons and in the peripheral nervous system

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