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A Badache

Publications and source records attributed to A Badache.

18 recordsLinked to original sources
PubMed2001

An essential role for Src kinase in ErbB receptor signaling through the MAPK pathway.

ErbB receptor tyrosine kinases are activated by multiple ligands such as epidermal growth factor (EGF) and neuregulins (NRGs), leading to stimulation of intracellular signaling pathways, including the mitogen-activated protein kinase (MAPK) cascade. We show here that Src kinase is essential for rapid EGF- and NRG-induced MAPK activation when the breast carcinoma cell lines T47D and SKBR3 are stimulated with low concentrations of ligand. In the presence of the pharmacological inhibitor CGP77675, which specifically blocks the activity of Src family kinases, ligand-induced MAPK activation was almost completely blocked at 5 min. Although this block was only transient, inactivation of Src suppressed ligand-induced transcription from a MAPK-responsive promoter. At the molecular level, the initial inhibition of MAPK by Src inactivation correlated with impaired ligand-induced Shc phosphorylation. Surprisingly, Src inhibition affected neither association of Shc with ErbB receptors nor phosphorylation of receptor-bound Shc. Thus, ErbB signaling requires the engagement of a novel Src-dependent route to MAPK, to trigger its rapid activation and subsequent efficient stimulation of transcription.

Adaptor Proteins, Signal Transducing↗
PubMed2001

Interleukin 6 inhibits proliferation and, in cooperation with an epidermal growth factor receptor autocrine loop, increases migration of T47D breast cancer cells.

Interleukin (IL)-6, a multifunctional regulator of immune response, hematopoiesis, and acute phase reactions, has also been shown to regulate cancer cell proliferation. We have investigated IL-6 signaling pathways and cellular responses in the T47D breast carcinoma cell line. The IL-6-type cytokines, IL-6 and oncostatin M, simultaneously inhibited cell proliferation and increased cell migration. In T47D cells, IL-6 stimulated the activation of Janus-activated kinase 1 tyrosine kinase and signal transducers and activators of transcription (STAT) 1 and STAT3 transcription factors. Expression of dominant negative STAT3 in the cells strongly reduced IL-6-mediated growth inhibition but did not prevent IL-6-induced cell migration. IL-6 treatment led to activation of the mitogen-activated protein kinase (MAPK) and the phosphatidylinositol 3'-kinase (PI3K) pathways. Inhibition of MAPK or PI3K activity reversed IL-6- and oncostatin M-stimulated migration. Because cross-talk between cytokine receptors and members of the ErbB family of receptor tyrosine kinases has been described previously, we have examined their interaction in T47D cells. Down-regulation of ErbB receptor activity, through the use of specific pharmacological inhibitors or dominant negative receptor constructs, revealed that IL-6-induced MAPK activation was largely dependent on epidermal growth factor (EGF) receptor activity, but not on ErbB-2 activity. Using a monoclonal antibody that interferes with EGF receptor-ligand interaction, we have shown that in T47D cells, IL-6 cooperates with an EGF receptor autocrine activity loop for signaling through the MAPK and PI3K pathways and for cell migration. Both the tyrosine phosphatase SHP-2 and the multisubstrate docking molecule Gab1, which are potential links between IL-6 and the MAPK/PI3K pathways, were constitutively associated with the active EGF receptor. On IL-6 stimulation, SHP-2 and Gab1 were recruited to the gp130 subunit of the IL-6 receptor and tyrosine phosphorylated, allowing downstream signaling to the MAPK and PI3K pathways. Thus, in T47D breast carcinoma cells, IL-6 acts in synergy with EGF receptor autocrine activity to signal through the MAPK/PI3K pathways. Cooperation between IL-6 and the EGF receptor in T47D breast carcinoma cells illustrates how a combination of multiple stimuli, either exogenous or endogenous, may result in synergistic cellular responses.

Adaptor Proteins, Signal Transducing↗
PubMed2001

The ErbB receptor tyrosine family as signal integrators.

ErbB receptor tyrosine kinases (RTKs) and their ligands have important roles in normal development and in human cancer. Among the ErbB receptors only ErbB2 has no direct ligand; however, ErbB2 acts as a co-receptor for the other family members, promoting high affinity ligand binding and enhancement of ligand-induced biological responses. These characteristics demonstrate the central role of ErbB2 in the receptor family, which likely explains why it is involved in the development of many human malignancies, including breast cancer. ErbB RTKs also function as signal integrators, cross-regulating different classes of membrane receptors including receptors of the cytokine family. Cross-regulation of ErbB RTKs and cytokines receptors represents another mechanism for controlling and enhancing tumor cell proliferation.

Female↗
PubMed2000

Myelin basic protein (MBP) and MBP peptides are mitogens for cultured astrocytes.

After CNS demyelination, astrogliosis interferes with axonal regeneration and remyelination. We now provide evidence that myelin basic protein (MBP) can contribute to this observed astrocyte proliferation. We found that astrocytes grown in either serum-containing or serum-free medium proliferate in response to MBP. The mitogenic regions of MBP in both media were MBP(1-44), MBP(88-151) and MBP(152-167). The mitogenic effect of these individual peptides was potentiated by simultaneous treatment with microglia conditioned media (CM). MBP-induced proliferation was inhibited by suramin at concentrations known to block the fibroblast growth factor receptor (FGFR), whereas neither MBP(1-44), MBP(88-151) nor MBP(152-167) were affected. Cholera toxin B, that binds to ganglioside GM(1), inhibited the mitogenicity of MBP(1-44) and had no significant effect on the mitogenicity of MBP, MBP(88-151) or MBP(152-167). Treatment of astrocytes with MBP and MBP(152-167) caused a modest and transitory elevation of intracellular calcium, whereas treatment with MBP(1-44) resulted in a substantial and sustained increase in intracellular calcium. These results suggest that for cultured astrocytes 1) FGFR and extracellular calcium play a major role in MBP mitogenicity; 2) MBP(1-44), MBP(88-151) and MBP(152-167) are the mitogenic regions of MBP; 3) MBP(1-44) and MBP(152-167) interact with ganglioside GM(1) and FGFR, respectively; 4) Component(s) present in microglial CM potentiate the mitogenicity of MBP(1-44), MBP(88-151) and MBP(152-167). These data support the hypothesis that MBP related peptides in conjunction with microglial secreted factors may stimulate astrogliosis after demyelination in vivo.

Animals↗
PubMed1999

Phosphorylation of CREB in axon-induced Schwann cell proliferation.

Axonal contact regulates Schwann cell (SC) proliferation during development. However, the intracellular signal transduction pathways involved in the axon-induced proliferation of SC have not been described. We have previously shown that SC proliferation induced by axolemma-enriched fractions (AEF) is accompanied by increased expression of cyclic AMP-responsive element binding protein, CREB. We now report the AEF and dorsal root ganglion neuritic-induced signal transduction pathway(s) which regulate the phosphorylation of CREB that correlate with the SC proliferative response. The phosphorylated form of CREB was significantly increased after 16 hr of axonal stimulation, continued to increase for 48 hr, and subsequently decreased as monitored by immunocytochemistry and Western blot analysis. Treatment with protein kinase A (PKA) inhibitor, H89, completely abolished both the CREB activation and SC proliferation. In contrast, treatment with protein kinase C (PKC) inhibitor (bisindolylmaleimide) inhibited AEF-induced SC proliferation, but did not immediately affect CREB phosphorylation. These data are consistent with the view that PKA and PKC pathways are essential for AEF-induced SC proliferation. Since PKC can influence SC proliferation without initially affecting CREB phosphorylation, PKC may regulate SC proliferation at pathways distal to the immediate CREB activation.

Animals↗
PubMed1998

Expression of Kit in neurofibromin-deficient human Schwann cells: role in Schwann cell hyperplasia associated with type 1 neurofibromatosis.

Type 1 Neurofibromatosis (NF1) is characterized by the formation of neurofibromas, benign tumors composed mainly of Schwann cells, which can turn malignant to form neurofibrosarcomas. Neurofibromin, the protein product of the Nf1 gene, is believed to act as a tumor suppressor, accelerating the conversion of the oncogene Ras to its inactive form. The absence of neurofibromin could therefore lead to higher Ras activity in Schwann cells, resulting in uncontrolled growth through a cascade of events not yet elucidated. We describe the abnormal expression of high levels of the Kit tyrosine kinase receptor in both NF1-derived Schwann cell lines and tissue, as compared to primary Schwann cells or schwannoma-derived cells. High levels of Kit expression in the neurofibrosarcoma-derived Schwann cells correlate with a decrease in neurofibromin expression. Using inhibitors of tyrosine kinase receptors, we found that proliferation of the neurofibrosarcoma-derived cells is dependent upon activation of a subclass of tyrosine-kinase receptors. The proliferation of these cells is not dependent upon an autocrine loop involving typical Schwann cell mitogens. Finally, the proliferation of the neurofibrosarcoma-derived Schwann cells can be increased by stimulation with Kit ligand. These data implicate Kit as one of the components leading to the Schwann cell hyperplasia observed in NF1.

Animals↗
PubMed1998

Neurofibrosarcoma-derived Schwann cells overexpress platelet-derived growth factor (PDGF) receptors and are induced to proliferate by PDGF BB.

Neurofibromatosis type 1 (NF1) is characterized by the formation of neurofibromas, benign tumors of the peripheral nerve consisting essentially of Schwann cells, which can sometimes turn malignant to form neurofibrosarcomas. The mechanism of progression toward a malignant phenotype remains largely unknown. In this report, we show that platelet-derived growth factor (PDGF) BB, and to a lesser extent fibroblast growth factor 2, are mitogenic for two neurofibrosarcoma-derived Schwann cell lines, but not for a Schwann cell line derived from a schwannoma (from a non-NF1 patient) or for transformed rat Schwann cells. Levels of expression of both PDGF receptor alpha and beta are significantly increased in the two neurofibrosarcoma-derived cell lines compared to the non-NF1 Schwann cell lines. The level of tyrosyl-phosphorylated PDGF receptor beta is strongly increased upon stimulation by PDGF BB. In comparison, only modest levels of tyrosyl-phosphorylated PDGF receptor alpha are observed, upon stimulation by PDGF AA or PDGF BB. Accordingly, PDGF AA is only a weak mitogen for the neurofibrosarcoma-derived cells by comparison to PDGF BB. These results indicate that the mitogenic effect of PDGF BB for the neurofibrosarcoma-derived Schwann cell lines is primarily transduced by PDGF receptor beta. Neu differentiation factor beta, a potent mitogen for normal Schwann cells, was unable to stimulate proliferation of the transformed Schwann cell lines, due to a dramatic down-regulation of the erbB3 receptor. Therefore, aberrant expression of growth factor receptors by Schwann cells, such as the PDGF receptors, could represent an important step in the process leading to Schwann cell hyperplasia in NF1.

Animals↗
PubMed1995

Human lymphocyte activation is associated with the early and high-level expression of the endogenous lectin CSL at the cell surface.

Lymphocytes undergo activation in response to antigens, cytokines, lectins and antibodies interacting with specific cell-surface molecules or through substances influencing signal transduction pathways. This study shows that human T- and B-cells stimulated using phorbol esters or plant lectins express early (2 h using phorbol esters and 24 h using plant lectins) a high level of a polyvalent carbohydrate-binding protein, the cerebellar soluble lectin (CSL), which is in part externalized. The lectin, immunologically related to CDw70, interacts with specific glycoprotein ligands of the lymphocyte surface, including CD3 on T-cells and CD24 on B-cells. Major changes in phosphorylations associated with activation appear as largely CSL-dependent since they are specifically inhibited by anti-CSL Fab fragments. It is suggested that the lectin induces the clustering of specific cell-surface glycoproteins and plays the role of an endogenous amplifier of activation signals.

Agglutination Tests↗
PubMed1995

An endogenous lectin and its glycoprotein ligands are triggering basal and axon-induced Schwann cell proliferation.

The proliferation of Schwann cells (the myelinating cells of the peripheral nervous system) is stimulated by the contact with axonal membranes. It is suggested that the endogenous carbohydrate-binding protein (lectin) cerebellar soluble lectin (CSL) bound to ligands at the surface of axonal preparations is mitogenic for Schwann cells. Both autocrine and axon-stimulated Schwann cell proliferations seem to be dependent on the presence of CSL and its ligands at the Schwann cell surface, as suggested by the effects of N-glycosylation inhibitors and anti-CSL Fab fragments. These data suggest that CSL regulates Schwann cell proliferation by clustering of a few glycoprotein ligands at the cell surface, consequently modulating phosphorylations.

1-Deoxynojirimycin↗
PubMed1994

Carbohydrates and soluble lectins in the regulation of cell adhesion and proliferation.

There is a large body of suggestions that complex carbohydrates play a role in the regulation of cell adhesion and cell proliferation. Many reports have emphasized that proteoglycans, glycoproteins or glycolipids are participating to cell adhesion mechanisms. The use of polyvalent anti-carbohydrate antibodies and plant lectins as well as the use of glycosylation inhibitors suggested that cell proliferation can be modulated by surface carbohydrates. The dating experiment of Burger and Noonan (1970) showing restoration of contact inhibition of malignant cells by monovalent concanavalin A was a determining experiment. However, in the latter as in the others, no precise mechanism was demonstrated how carbohydrates can be involved in adhesion and proliferation. New insights were opened with the discovery of vertebrate membrane-bound and soluble lectins. The latter generally display agglutinating activities in in vitro systems, suggesting that they were potential cell adhesion molecules, by forming bridges between cell surface carbohydrates. These polyvalent molecules may be also considered as clustering agents for their cell surface ligands, consequently generating signals for cell proliferation and/or differentiation.

Animals↗
PubMed1993

Cerebellar soluble lectin and its glycoprotein ligands in the developing brain of control and dysmyelinating mutant mice.

The levels of an endogenous lectin, the cerebellar soluble lectin (CSL) and of its endogenous glycoprotein ligands were studied using immunoblotting and affinoblotting techniques in the forebrain of quaking, shiverer and jimpy dysmyelinating mutant mice and their respective control littermates during the postnatal development. In the controls of the mutant mice, the level of CSL showed an important increase between days 5-18 then a stabilization, although at all ages the level of CSL was reduced (at least 15%) in the control littermate of the shiverer mutant. In the shiverer mutant the developmental pattern is similar to the control but was reduced by 50% as compared to the control. In the jimpy mutant an erratic development of CSL was observed which was with quasi absence of CSL at days 12 and 25. Variation of CSL levels in the quaking brain were also observed. CSL glycoprotein ligands also showed variable developmental profiles with a special persistence with ageing of CSL-binding glycoproteins in the quaking and jimpy mice. Developmental variations were also observed between the different control littermates. These results are discussed in view of developmental roles attributed to CSL and its glycoproteins ligands in cell adhesion mechanism during brain ontogenesis and especially myelination.

Aging↗
PubMed1993

Lesion-induced re-expression of neonatal recognition molecules in adult rat cerebellum.

It has been previously shown that sectioning of parallel fibers in the cerebellar molecular layer of adult rats gave rise to rapid reinnervation of the target cells, i.e., Purkinje cells. This paper reports that such a reinnervation is accompanied by reexpression (partial and total) of two developmentally regulated complementary molecules. These are an endogenous mannose-binding lectin, called R1, which reappears at the surface of the dendrites of Purkinje cells, and an endogenous glycoprotein ligand of R1, the 31 kDa glycoprotein, which seems to be neosynthetized and transported to the surface of parallel fibers. In this system, embryonic N-CAM is not reexpressed in neurons but reappears in reactive astrocytes in the vicinity of the lesion. The reexpression of recognition molecules (lectin and glycoprotein ligand) involved in normal synaptogenesis, may constitute the molecular basis for repair of nervous circuits in the adult as well.

Animals↗
PubMed1992

Glycoproteins and lectins in cell adhesion and cell recognition processes.

The discovery of endogenous lectins having specific and high affinity for the carbohydrate portions of glycoproteins has opened up new directions in the field of cell adhesion and cell recognition. Two endogenous lectins, termed as CSL and R1, initially isolated from the rat cerebellum and having a wide distribution in mammalian tissues, have been shown to participate in essential mechanisms of cell adhesion. The membrane-bound lectin R1 seems to be involved in transient recognition between neuronal cells, followed by elimination of the glycoprotein ligands at the surface of the recognized cell. In contrast, CSL is a molecule involved in adhesion between various normal or transformed cells since it participates in the formation of tight junctions. The glycoprotein ligands recognized with higher affinity by these two lectins seem to possess a special structure which defines a sub-class of oncofetal HNK-1 glycans. The over-expression of the glycoprotein ligands of these lectins in most transformed cells provides new tools for understanding the underlying mechanism of malignant transformation as well as the generation of signals through cell adhesion.

Animals↗
PubMed1992

Carbohydrate moieties of myelin-associated glycoprotein, major glycoprotein of the peripheral nervous system myelin and other myelin glycoproteins potentially involved in cell adhesion.

The myelin-associated glycoprotein (MAG) and the major glycoprotein of the peripheral nervous system myelin (P0) are two members of the family of cell adhesion molecules (CAMs). A role in cell adhesion of the carbohydrate moiety of these molecules has been attributed to the presence of N-glycans bearing the HNK-1 carbohydrate epitope. On the other hand, it has been suggested that these glycoproteins could be ligands of an endogenous mannose-binding lectin present in myelin, the cerebellar soluble lectin (CSL). In order to further document the heterogeneity of the glycans of these two CAMs, we have used several probes: an anti-carbohydrate antibody of the HNK-1 type, called Elec-39, the plant lectin concanavalin A (ConA), and the endogenous lectin CSL involved in myelin compaction. This study shows that CSL binds to a small proportion of the polypeptide chains of MAG found in adult CNS of rats and man and the polypeptide chains of P0 molecules from adult human and rat sciatic nerve. For MAG from adult rat brain, the binding of CSL is restricted to glycans of polypeptide chains which could be separated from the others according to their solubility properties. These MAG molecular entities react also with the Elec-39 antibody and with ConA. These results confirm that P0 and MAG are heterogeneous in their carbohydrate moieties.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

The endogenous lectin cerebellar soluble lectin and its ligands in central nervous system myelin of myelin-deficient (mld) mutant mice.

The myelin-deficient (mld) mutation is autosomal recessive mutation in the murine CNS exhibiting severe hypomyelination. The primary defect results in a drastic reduction of myelin basic protein synthesis caused by a duplication of the myelin basic protein gene with partial inversion of the upstream gene copy. The severe deficit of myelin basic protein is responsible for the absence of the major dense line but cannot explain the heterogeneity of myelin compaction found in mld. We have tested the hypothesis that the endogenous cerebellar soluble lectin (CSL) and/or its endogenous glycoprotein ligands could be involved in myelin abnormalities in the dysmyelinating mutant, mld. Immunocytochemical and immunoblotting techniques showed that the CSL level was not reduced significantly in the mld mutant. Furthermore, two ligands of CSL, the myelin-associated glycoprotein and an axonal glycoprotein, with a relative molecular mass of 31 kDa, were not decreased in level in the purified myelin fraction isolated from mld mice. In contrast, three minor glycoprotein ligands of CSL of relative molecular mass of 23, 18, and 16 kDa were greatly reduced in content. The reduced concentration of these low-molecular-mass glycoproteins in mld myelin suggests that they are constituents of compact myelin. Furthermore, the observation that CSL is specifically localized in vivo in regions where mld myelin is more compact and absent from regions devoid of myelin compaction may suggest that the endogenous CSL lectin, as well as its minor glycoprotein ligands, plays a role in the stabilization of the myelin sheath.

Animals↗
PubMed1991

Involvement of the endogenous lectin CSL in adhesion of Chinese hamster ovary cells.

Immunochemical localization of an endogenous mannose-binding protein, the cerebellar soluble lectin (CSL; Zanetta et al., J. Neurochem. 49, 1250-1257 (1987)), in Chinese hamster ovary cells indicated its high concentration in areas of contact between cells. This suggested its role in cell adhesion. The pattern of staining differed significantly in the cells cultured in suspension from that grown as monolayer. In cells maintained for a short time as suspension, the extracellular CSL immunoreactivity was found mainly in close apposition to the plasma membrane including contact areas. In cells cultured as monolayer, extracellularly, the lectin was found both at the cell surface and in a 75-nm thick layer between two cells, apparently adhering to the cell surface through bridges. Endogenous glycoprotein ligands of CSL were present in the cultures of CHO cells, both as membrane-bound glycoproteins and as glycoprotein ligands soluble in the presence of mannose in the absence of detergent. The lectin CSL induced adhesion between these cells as evident by low concentration of anti-CSL Fab fragments inhibiting such adhesion. These data suggested that adhesion between CHO cells occurs, in part, through a glycobiological recognition system involving CSL. This mechanism should be taken into account for the interpretation of experiments of transfection in CHO cells of the genes of glycoproteins involved in cell adhesion.

Animals↗
PubMed1990

Endogenous cerebellar soluble lectin and its ligands in central nervous system myelin of quaking and jimpy mutant mice.

The presence of an endogenous 'cerebellar soluble lectin' (CSL) involved in myelin compaction and myelination was analyzed in the dysmyelinating mutant mice quaking and jimpy. The primary defect in these mutations with severe hypomyelination is still unknown in the quaking mutant but results from a single mutation in the proteolipid protein gene in the jimpy mutant. Both immunocytochemical and immunoblotting techniques showed that CSL was not considerably reduced in its expression in the myelin fraction purified from adult quaking mutants. Furthermore, the myelin-associated glycoprotein and an axonal glycoprotein with a relative molecular weight (Mr) of 31 kilodaltons (kDa) were not decreased in quaking mice. This contrasted with several glycoproteins of Mr 23, 18, 16 and 12 kDa which were absent from the purified quaking myelin. In myelin preparations obtained from the jimpy mutant the CSL level was considerably reduced. This defect did not result from a deficient synthesis of CSL. However, as in the quaking mutation low-Mr glycoproteins were lacking. The nature of the low-Mr glycoproteins absent in quaking and jimpy mice is discussed in relation to previous reports on myelin glycoproteins. In the various mutants, due to different primary mutations, a similar absence of myelin compaction was observed, which could be associated with a deficient level of low-Mr glycoproteins. It is thus postulated that these molecules are essential for ensuring myelin compaction as ligands for the endogenous CSL.

Animals↗