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T Maciag

Publications and source records attributed to T Maciag.

At least 55 records · Page 3Linked to original sources
PubMed1995

The release of fibroblast growth factor-1 from NIH 3T3 cells in response to temperature involves the function of cysteine residues.

Fibroblast growth factor (FGF)-1 is released from NIH 3T3 cells in response to heat shock as a biologically inactive protein that is unable to bind heparin and requires activation by (NH4)2SO4 to generate a biologically active extracellular heparin-binding growth factor (Jackson, A., Friedman, S., Zhan, X., Engleka, K. A., Forough, R., and Maciag, T. (1992) Proc. Natl. Acad. Sci. USA 89, 10691-10695). To further study the mechanism of FGF-1 release in response to heat shock (42 degrees C), we examined the kinetics of FGF-1 release from FGF-1-transfected NIH 3T3 cells and observed that the cells require at least 1 h of exposure to heat shock conditions for the release of FGF-1. Interestingly, agents that interfere with the function of the endoplasmic reticulum-Golgi apparatus, exocytosis, and the multidrug resistance pathway (brefelden A, methylamine, and verapamil, respectively) do not inhibit the release of FGF-1 in response to temperature; rather, they exaggerate the release of FGF-1. Because immunoblot analysis of FGF-1 in the conditioned medium of heat-shocked NIH 3T3 cells revealed the presence of a minor band with an apparent molecular weight of a FGF-1 homodimer and because we have previously shown that FGF-1, but not FGF-2, is able to form a homodimer in response to chemical oxidation by CuCl2 (Engleka, K. A., and Maciag, T. (1992) J. Biol. Chem. 267, 11307-11315), we examined whether reducing agents would substitute for (NH4)2SO4 and activate extracellular FGF-1. Indeed, dithiothreitol and reduced glutathione are able to individually generate a FGF-1 monomer as a heparin-binding protein from the conditioned medium of heat-shocked NIH 3T3 cell transfectants. To confirm that cysteine residues are involved in the release of FGF-1 in response to temperature, we used mutagenesis to prepare a human FGF-1 Cys-free mutant in which Cys30, Cys97, and Cys131 were converted to serine. Analysis of the release of the FGF-1 Cys-free mutant in NIH 3T3 cells transfected with the FGF-1 Cys-free mutant demonstrated that the FGF-1 Cys-free mutant is not released into the conditioned medium in response to temperature. Interestingly, exposure of the NIH 3T3 cell FGF-1 Cys-free transfectants to brefelden A followed by heat shock also demonstrated the absence of the extracellular FGF-1 Cys-free mutant.(ABSTRACT TRUNCATED AT 400 WORDS)

3T3 Cells↗
PubMed1995

Molecular mechanisms of angiogenesis: fibroblast growth factor signal transduction.

The fibroblast growth factors are a family of structurally related polypeptides that are mitogenic for a broad range of cell types as well as mediators of a wide spectrum of developmental and pathophysiological processes in vivo and in vitro. The fibroblast growth factor family presently consists of nine distinct members. Indeed, the FGF prototypes FGF-1 (acidic) and FGF-2 (basic) are well described as modifiers of angiogenesis. The absence of a signal sequence to direct their secretion and their ability to traffic to the nucleus are unique structural features that may be relevant to the regulation of their activities. The FGF receptor family consists of four transmembrane receptor tyrosine kinases. Each of these receptors give rise to multiple isoforms as a result of alternative splicing of their mRNAs. The significance of these multiple isoforms is not fully understood; however it is known that alternative splicing in the extracellular domain of these receptors results in altered ligand binding specificities. In addition, alternative splicing in the cytoplasmic domain results in isoforms with increased oncogenic potential. This review will describe recent insights into the pathways used for the regulation of FGF secretion and cellular trafficking as well as signaling by FGFRs.

Alternative Splicing↗
PubMed1995

FGF-1 in normal and regenerating kidney: expression in mononuclear, interstitial, and regenerating epithelial cells.

The proximal tubule epithelium regenerates following nephrotoxic damage. To determine the role of fibroblast growth factors (FGFs) in the regeneration of rat proximal tubule epithelial (RPTE) cells, we investigated proliferation, differentiation, and FGF-1 expression in vivo in rat kidney before and after nephrotoxic damage to the proximal tubule epithelium caused by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine administration. In undamaged kidneys, FGF-1 was expressed in distal tubule elements, including cortical and medullary collecting ducts, as well as in blood vessels and glomeruli, but was absent in RPTE. One day after damage, there was an increase in proliferation of surviving proximal tubule epithelial cells and a coincident increase in FGF-1 expression in invading mononuclear cells. After this initial burst of proliferation, FGF-1 expression increased in poorly differentiated vimentin-positive regenerative epithelial cells, indicating that autocrine FGF-1 expression in the regenerative epithelium is a later event in the regeneration process. FGF-1 staining persisted in foci of macrophages, interstitial cells, and nephropathic tubules within areas of interstitial expansion 2 wk after damage. We concluded that transient paracrine and autocrine expression of FGF-1 could play mitogenic and/or morphogenic roles during tubular regeneration. Persistent expression in macrophages, fibroblasts, and nephropathic tubules may be associated with tubular degeneration. FGF-1 expression may be an important contributor to both tubular regeneration and degenerative disease following toxicant exposure.

Animals↗
PubMed1995

Extracellular FGF-1 acts as a lens differentiation factor in transgenic mice.

The vertebrate ocular lens undergoes a spatially defined pattern of differentiation which may be regulated by the ocular distribution of proteins from the fibroblast growth factor (FGF) family. The ability of altered FGF-1 (acidic FGF) distribution to disrupt the normal pattern of lens differentiation was evaluated by the production of transgenic mice which express FGF-1 under the control of the lens-specific alpha A-crystallin promoter. Since FGF-1 lacks a classical signal peptide consensus sequence, transgenic mice were also produced with a chimeric construct containing the signal peptide sequence of the FGF-4 gene fused in frame to the coding sequences of the FGF-1 cDNA in order to obtain extracellular expression of the transgene. The presence of transgenic mRNA and protein was confirmed by in situ hybridization, Western analysis and immunohistochemistry. The ocular histology of newborn and young adult transgenic mice expressing FGF-1 without a signal peptide appeared normal. In contrast, mice expressing secreted FGF-1 exhibited lens abnormalities including the elongation of anterior epithelial cells. Epithelial cell elongation was accompanied by expression of the fiber cell differentiation marker, beta-crystallin. These observations provide an in vivo demonstration that FGF-1 can induce anterior lens epithelial cells to express characteristics consistent with the onset of fiber cell differentiation. The transgenic induction of differentiation confirms that normal lens morphology reflects an asymmetric distribution of inductive factors within the eye.

Animals↗
PubMed1994

Intact and functional fibroblast growth factor (FGF) receptor-1 trafficks near the nucleus in response to FGF-1.

Exogenous fibroblast growth factor-1 (FGF-1) associates with the nucleus in a receptor-dependent manner during the entire G1 period of the BALB/c 3T3 cell cycle (Zhan, X., Hu, X., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 9611-9620). To further study the role of the FGF receptor (FGFR) during this translocation, the intracellular fate of FGFR-1 protein and enzymatic activity was examined. Immunoprecipitation using multiple FGFR-1 antibodies followed by an in vitro tyrosine kinase activity assay enabled us to identify FGFR-1 as a 130-kDa phosphotyrosine-containing protein associated with the nuclear fraction of NIH 3T3 cells exposed to FGF-1. While FGFR-1 tyrosine kinase activity could be detected as a nuclear-associated protein after a 2-h exposure of the NIH 3T3 cells to FGF-1, this activity appeared to be maximal in the nuclear fraction between 4 and 12 h after FGF-1 treatment. In addition, analysis by confocal immunofluorescence microscopy of quiescent and FGF-1-stimulated NIH 3T3 cells reveal a prominent perinuclear FGFR-1 staining pattern in the cells exposed to FGF-1 but not in the quiescent population. We also observed FGFR-1 associated with the nuclear fraction in FGFR-1-transfected L6 rat myoblasts, which are known to be refractive to exogenous FGF-1 and express relatively low levels of endogenous FGFR-1. In addition, these cells also exhibited the presence of a 145-kDa phosphoprotein in the nuclear fraction that was recognized by FGFR-1 antibodies. These results suggest that the FGFR-1 may be translocated near the nucleus upon interaction with its ligand during the entire G1 period of the NIH 3T3 cell cycle as a structurally intact and functional tyrosine kinase that may be accessible to perinuclear polypeptides as a regulatory enzyme.

3T3 Cells↗
PubMed1994

Association of fibroblast growth factor receptor-1 with c-Src correlates with association between c-Src and cortactin.

The initiation of maximal DNA synthesis by fibroblast growth factor (FGF)-1 requires the presence of the growth factor during the entire G0 to G1 transition period of the cell cycle (Zhan, X., Hu, X., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 9611-9620). During this time, the phosphorylation of several novel proteins on tyrosine residues occurs, and one of these phosphotyrosyl-containing proteins has been characterized as the murine homolog of the chicken cortactin gene (Zhan, X., Hu, X., Hampton, B., Burgess, W.H., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 24427-24431), a putative substrate for v-Src. We have examined the possibility that FGF-1 employs c-Src or Src-like kinases as signaling intermediates during the mid and late G1 phase of the NIH 3T3 cell cycle using immunoprecipitation and immunoblot analysis. We have demonstrated that c-Src can associate with cortactin in a FGF-1-dependent manner. We have also demonstrated that a monoclonal antibody prepared against FGF receptor (R)-1 is able to co-precipitate Src-related proteins in lysates from FGF-1-treated NIH 3T3 cells. Furthermore, a kinase-active form of FGFR-1 expressed in a bacterial system was also able to associate with Src kinases in a manner dependent on the phosphorylation status of the FGFR-1 protein. Lastly, the Src homology (SH)-2 domain of v-Src was able to recognize a recombinant form of FGFR-1. Because (i) the association between FGFR-1 and Src-like kinases exhibits kinetics similar to those observed between the Src kinases and cortactin and (ii) the Src-SH2 domain is likely to be involved in the association with FGFR-1, we propose that the association of c-Src with activated FGF receptors may be responsible for the tyrosine phosphorylation of cortactin during the mid to late G1 phase of the cell cycle.

3T3 Cells↗
PubMed1994

Induction of cyclooxygenase-2 by interleukin-1 alpha. Evidence for post-transcriptional regulation.

Prostanoids, produced by diverse cell types, modulate a variety of pathophysiological processes. The rate of prostanoid synthesis is determined, in part, by the levels of prostanoid-synthetic enzymes, such as cyclooxygenase (Cox), a rate-limiting enzyme in the conversion of arachidonic acid to prostanoids. While two Cox genes have been identified, Cox-2 is unique because it is highly induced in response to cell activation processes including inflammation. We have studied the effect of interleukin-1 alpha (IL-1 alpha), a proinflammatory cytokine that facilitates its actions in part by inducing the synthesis of prostanoids, on the expression of Cox-2 in a human cell line (ECV304) and demonstrated that IL-1 alpha induces a sustained increase in the expression of the Cox-2 mRNA as well as the functional enzyme. Three mechanistically distinct inhibitors of translation stimulated the expression of the Cox-2 mRNA and potentiated the effect of IL-1 alpha. Furthermore, IL-1 alpha induced rapid but transient activation of Cox-2 transcription and, in the absence of transcription, prolonged the half-life of the Cox-2 mRNA. Together, these data suggest that post-transcriptional mechanisms are important in the sustained induction of the Cox-2 mRNA and that IL-1 alpha may increase the stability of the Cox-2 transcript.

Base Sequence↗
PubMed1994

Mutagenesis of the nuclear localization sequence in EGF-1 alters protein stability but not mitogenic activity.

Fibroblast growth factor (FGF)-1 is able to translocate to the nucleus as an exogenous protein and a deletion of the nuclear localization sequence near the NH2-terminus of FGF-1 (FGF-1(28-154)) yields a recombinant polypeptide with impaired mitogenic activity. To study the significance within this region (NYKKPK), five FGF-1 point mutations were constructed and their recombinant protein forms analyzed. Interestingly, none of the mutant protein products showed a significant reduction in mitogenic activity when compared to wild-type protein. Because these data suggested possible structural instability within the FGF-1(28-154) deletion mutant, protein structure was examined by fluorescence spectroscopy. Indeed, the FGF-1 point mutations which had similar mitogenic activity to wild-type FGF-1 displayed similar thermal fluorescence spectroscopy patterns as wild-type protein, but FGF-1(28-154) exhibited altered fluorometric profiles. However, the mitogenic activity and structural stability of FGF-1(28-154) was dependent upon the method used to purify the recombinant, whereas purification methods did not effect FGF-1(21-154) mitogenic activity. These studies suggest that the reduced mitogenic activity observed in preparations of the FGF-1(28-154) deletion mutant may be the result of structural instability and fluorescence spectroscopy cannot be used to predict FGF-1 stability.

3T3 Cells↗
PubMed1994

Post-transcriptional regulation of interleukin 1 alpha in various strains of young and senescent human umbilical vein endothelial cells.

Human umbilical vein endothelial cell (HUVEC) senescence in vitro is characterized by the loss of proliferative potential and an increase in cell size. Because HUVEC senescence in one strain (H101) has been characterized by the increase in the steady-state mRNA level for the signal-peptideless cytokine, interleukin (IL) 1 alpha, we have examined young and senescent populations of five additional HUVEC strains (H3605, H103, H928, H929, and H930) to determine whether the elevated levels of IL-1 alpha mRNA could be observed in all HUVEC strains. Consistent with the data from strain H101, strains H3605 and H930 also exhibited a low steady-state level of the IL-1 alpha mRNA in young populations compared to elevated levels of IL-1 alpha mRNA in the senescent populations. However, three strains (H103, H928, and H929) did not exhibit reduced levels of IL-1 alpha mRNA in the young populations, and interestingly, strain H928, at times, expressed relatively high IL-1 alpha mRNA levels in the young populations. In addition, expression of the steady-state level of plasminogen activator inhibitor 1 and cyclooxygenase 2 was elevated in senescent populations of all HUVEC strains examined, whereas young populations exhibited a low level of expression for these genes regardless of the IL-1 alpha mRNA level. Further, the level of the IL-1 alpha polypeptide was elevated in senescent HUVEC populations relative to young populations that expressed either a high or low level of the IL-1 alpha mRNA. We have also demonstrated that the elevated level of IL-1 alpha mRNA in the senescent population of strain H3605 may be regulated by mRNA stability; however, this mechanism does not apply to all the HUVEC strains examined in this study. Thus, we suggest that while mRNA levels of the IL-1-response genes for plasminogen activator inhibitor 1 and cyclooxygenase 2 are appropriate markers for HUVEC senescence, HUVEC strain-specific post-transcriptional mechanisms may exist to regulate the function of IL-1 alpha as a modifier of HUVEC senescence in vitro.

Base Sequence↗
PubMed1993

Murine cortactin is phosphorylated in response to fibroblast growth factor-1 on tyrosine residues late in the G1 phase of the BALB/c 3T3 cell cycle.

We have previously reported that BALB/c 3T3 cells require a prolonged exposure to fibroblast growth factor (FGF)-1 for the stimulation of maximal DNA synthesis, and this event correlates with the tyrosine phosphorylation of novel proteins late in G1 including a protein termed p80/p85 (Zhan, X., Hu, X., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 9611-9620). We have purified, sequenced, and cloned the cDNA encoding p80/p85 and report that it is the murine homolog of the chicken cortactin gene and a member of the human hematopoietic specific-1 gene family. Immunochemical analysis of m-cortactin-tyrosine phosphorylation in response to FGF-1 demonstrates a biphasic phosphorylation pattern both as a weak immediate-early and strong mid to late G1 response protein. Because the chicken cortactin gene was originally isolated as a substrate for v-Src, FGF-1 may influence the enzymatic activity of other cell-associated tyrosine kinases which utilize p80/p85 (cortactin) as a polypeptide substrate.

3T3 Cells↗
PubMed1993

Identification of a nuclear localization sequence within the structure of the human interleukin-1 alpha precursor.

Recent studies have suggested that the signal peptide-less cytokine, interleukin (IL)-1 alpha, may play a role as an intracellular regulator of human endothelial cell proliferation in vitro (Garfinkel, S., Haines, D. S., Brown, S., Wessendorf, J., Gillespie, D. H., and Maciag, T. (1992) J. Biol. Chem. 267, 24375-24378). In order to determine the intracellular locale of the IL-1 alpha precursor, we fused the open reading frame of the IL-1 alpha precursor to the reporter gene beta-galactosidase (Gal) and studied the cellular distribution of the chimera in NIH 3T3 cells after transfection. Immunological and enzymatic analysis demonstrated that the IL-1 alpha:beta-Gal fusion protein was associated with the nucleus. To further define the region responsible for this activity, we ligated the mature form of IL-1 alpha (IL-1 alpha 113-271) and the IL-1 alpha precursor domain (IL-1 alpha 1-112) to beta-Gal. Analysis of the intracellular distribution of these chimeric polypeptides following transfection demonstrated a differential distribution of IL-1 alpha 1-112:beta-Gal in the nucleus and IL-1 alpha 113-271:beta-Gal in the cytosol. Because the IL-1 alpha precursor domain contains a sequence that resembles a nuclear translocation signal (KVLKKRRL, residues 79-86), we prepared an IL-1 alpha precursor point mutant in which Lys82 was replaced by Glu. Transfection of NIH 3T3 cells with the IL-1 alpha precursor point mutant (IL-1 alpha 1-271 Glu82:beta-Gal) resulted in a significant reduction in the ability of the IL-1 alpha precursor to associate with the nucleus and similar data were obtained as a result of Lys82 mutagenesis in the IL-1 alpha precursor domain (IL-1 alpha 1-112 Glu82:beta-Gal). These data suggest that the IL-1 alpha precursor contains a functional nuclear localization sequence within the structure of the precursor domain and Lys82 is critical for its function.

3T3 Cells↗
PubMed1993

A role for fibroblast growth factor type-1 in nephrogenic repair. Autocrine expression in rat kidney proximal tubule epithelial cells in vitro and in the regenerating epithelium following nephrotoxic damage by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine in vivo.

The regenerating proximal tubule epithelium in the rat has striking similarity with rat kidney proximal tubule epithelial cells (RPTE) in primary culture (Wallin, A., Zhang, G., Jones, T. W., Jaken, S., and Stevens, J. L. (1992) Lab. Invest. 66, 474-484). We used this in vitro model to investigate mechanisms which may regulate aspects of nephrogenic repair in vivo, and in particular the role of fibroblast (heparin-binding) growth factor type-1 (FGF-1) expression. FGF-1 was present predominantly as a 14.3-kDa polypeptide in rat kidney. Biological activity and mRNA for FGF-1 increased dramatically in primary RPTE in culture along with an increase in a 18.3-kDa FGF-1. Cycloheximide blocked FGF-1 expression in primary culture indicating that the increase represents newly synthesized factor rather than release from the extracellular matrix. The maximal increase in expression occurs after the peak of RPTE proliferation. Activity was not present in the medium but intracellular FGF-1 was released from RPTE by scrape wounding. Immunohistochemical analysis showed that FGF-1 expression increased in regenerating proximal tubule epithelial cells 5 days after nephrotoxic damage. Collectively, the data suggest that autocrine expression of FGF-1 by regenerating proximal tubule epithelial cells plays a role in the regulation of nephrogenic repair.

Animals↗
PubMed1993

Long term growth factor exposure and differential tyrosine phosphorylation are required for DNA synthesis in BALB/c 3T3 cells.

The importance of growth factor-mediated immediate-early cellular events to the cell cycle has influenced the development and identity of oncogenes and tumor suppressor genes as well as the concept that growth factors commit mammalian cells to enter a biochemical program that ultimately yields DNA synthesis. However, the mid and late events involved in the regulation of growth factor-induced signal transduction remain largely unknown. In this report we demonstrate that BALB/c 3T3 cells require continuous exposure to fibroblast growth factor (FGF)-1 for a minimum of 12 h to achieve near maximal DNA synthesis. This correlates with the continuous internalization of radiolabeled FGF-1 into the cytosol and nucleus of BALB/c 3T3 cells and the maintenance of a low level of FGF receptors on the cell surface during the entire G1 phase of the cell cycle. Further analysis demonstrates the maintenance of a continuous series of differential FGF-1-induced tyrosine phosphorylation events including the phosphorylation of phospholipase C-gamma as well as novel FGF receptor polypeptide substrates, p60, p85, p90, and p130 throughout the G1 phase of the BALB/c 3T3 cell cycle. The tyrosine phosphorylation events are biphasic during the 12-h period after the administration of FGF-1, and the second phase is characterized by hyper-tyrosine phosphorylation of p60, p85, and p130. Interestingly, NIH 3T3 cells which overexpress the FGF receptor-1 polypeptide demonstrate exaggerated tyrosine phosphorylation of p60 and p85 but not p90 and exhibit growth factor-independent cell proliferation. These results suggest that the initiation of DNA synthesis in BALB/c 3T3 cells by FGF-1 is regulated by a complex biochemical program that involves the continuous tyrosine phosphorylation of known and novel polypeptides throughout the G0 to G1 transition period of the cell cycle.

3T3 Cells↗
PubMed1993

Recombinant fibroblast growth factor-1 promotes intimal hyperplasia and angiogenesis in arteries in vivo.

The prototype members of the heparin-binding fibroblast growth factor (FGF) family, acidic FGF (FGF-1) and basic FGF (FGF-2), are among the growth factors that act directly on vascular cells to induce endothelial cell growth and angiogenesis. In vivo, the role of the FGF prototypes in vascular pathology has been difficult to determine. We report here the introduction, by direct gene transfer into porcine arteries, of a eukaryotic expression vector encoding a secreted form of FGF-1. This somatic transgenic model defines gene function in the arterial wall in vivo. FGF-1 expression induced intimal thickening in porcine arteries 21 days after gene transfer, in contrast to control arteries transduced with an Escherichia coli beta-galactosidase gene. Where there was substantial intimal hyperplasia, neocapillary formation was detected in the expanded intima. These findings suggest that FGF-1 induces intimal hyperplasia in the arterial wall in vivo and, through its ability to stimulate angiogenesis in the neointima, FGF-1 could stimulate neovascularization of atherosclerotic plaques. Potentially, gene transfer of FGF-1 could also be used as a genetic intervention to improve blood flow to ischaemic tissues in selected clinical settings.

Animals↗
PubMed1993

Differential transforming abilities of non-secreted and secreted forms of human fibroblast growth factor-1.

Fibroblast growth factor (FGF)-1(1-154), the precursor for acidic FGF-1(21-154), is a potent angiogenic polypeptide, the structure of which lacks a signal peptide sequence for secretion. To investigate the biological significance of this structural feature, we have attempted forced secretion of FGF-1 through fusion of the entire FGF-1 coding frame with the signal peptide (sp) from the hst/KS3 gene, a secretory member of the heparin-binding growth factor family. We also studied the transforming ability of the signal-less forms of FGF-1 comprising FGF(1-154) and FGF-1(21-154). The presence of a soluble and biologically active form of FGF-1 was readily detected in the conditioned medium of NIH 3T3 cells transfected with sp-hst/KS3:FGF-1(1-154) as demonstrated by Western blot analysis and DNA synthesis assays, whereas sp-hst/KS3:FGF-1(21-154) was not detectable in conditioned medium even though the protein was detected in cellular extracts. The secreted form of sp-hst/KS3:FGF-1(1-154) stimulated the proliferation of human umbilical vein endothelial cells in vitro and was able to induce receptor-mediated tyrosine phosphorylation. Furthermore, the forced secretion of biologically active FGF-1 resulted in NIH 3T3 cell transformation as demonstrated by altered morphology in vitro, the formation of discrete colonies in soft agarose, growth under serum-free conditions, and ability to rapidly form highly vascular tumors in vivo. Interestingly, sp-hst/KS3:FGF-1(21-154) also mediated the transition to a transformed phenotype despite the inability to detect extracellular FGF-1 in the media conditioned by these NIH 3T3 cell transfectants. Although the transfection of FGF-1(21-154) yielded similar NIH 3T3 cell morphologic changes, these transfectants did not grow under serum-free conditions or yield colonies in soft agarose, and formed tumors in vivo with delayed kinetics. Furthermore, the FGF-1(1-154) NIH 3T3 cell transfectants did not exhibit morphologic changes, and this may be due to the inability of mRNA to express protein. These data suggest that although non-sp forms of FGF-1 may alter the monolayer phenotype of NIH 3T3 cells in vitro, the ability of FGF-1 to transform NIH 3T3 cells requires the function of a sp-directed secretory pathway and suggests that this pathway increases tumorigenicity in vivo.

3T3 Cells↗
PubMed1993

Coexpression of phosphotyrosine-containing proteins, platelet-derived growth factor-B, and fibroblast growth factor-1 in situ in synovial tissues of patients with rheumatoid arthritis and Lewis rats with adjuvant or streptococcal cell wall arthritis.

Fibroblast growth factor (FGF)-1 and PDGF-B-like factors have been implicated in the pathobiology of RA and animal models of this disease. Since the receptors for FGF-1 and PDGF are tyrosine kinases, we examined the expression of tyrosine phosphorylated proteins (phosphotyrosine, P-Tyr) in synovial tissues from patients with RA and osteoarthritis (OA), and rats with streptococcal cell wall (SCW) and adjuvant arthritis (AA). Synovia from patients with RA and LEW/N rats with SCW and AA arthritis, in contrast to controls, stained intensely with anti-P-Tyr antibody. The staining colocalized with PDGF-B and FGF-1 staining. Comparative immunoblot analysis showed markedly enhanced expression of a 45-kD P-Tyr protein in the inflamed synovia. Treatment with physiological concentrations of dexamethasone suppressed both arthritis and P-Tyr expression in AA. P-Tyr was only transiently expressed in athymic nude Lewis rats and was not detected in relatively arthritis-resistant F344/N rats. These data suggest that (a) FGF-1 and PDGF-B-like factors are upregulated and may induce tyrosine phosphorylation of proteins in vivo in inflammatory joint diseases, (b) persistent high level P-Tyr expression is T lymphocyte dependent, correlates with disease severity, and is strain dependent in rats, (c) corticosteroids, in physiological concentrations, downregulate P-Tyr expression in these lesions.

Animals↗