Fibroblast Growth Factor-1 Induces Phosphofructokinase, Fatty Acid Synthase and Ca2+-ATPase mRNA Expression in NIH 3T3 Cells

Hsu, Debbie K.W.; Donohue, Patrick J.; Alberts, Gregory F.; Winkles, Jeffrey A.

doi:10.1006/bbrc.1993.2644

Cited by 41 publications

(21 citation statements)

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“…RNA (1 g) from quiescent or FGF-1/cycloheximide-treated NIH 3T3 cells was converted to cDNA using Moloney murine leukemia virus reverse transcriptase (Life Technologies, Inc.) and random hexamer primers (Roche Molecular Biochemicals) as described (16). PCR assays were performed using a degenerate sense protein kinase domain oligonucleotide primer (12) and a degenerate antisense leucine zipper domain oligonucleotide primer (17). An equivalent aliquot of each amplification mixture was subjected to electrophoresis in a 2% agarose gel, and DNA was visualized by ethidium bromide staining.…”

Section: Methodsmentioning

confidence: 99%

The Mitogen-inducible Fn14 Gene Encodes a Type I Transmembrane Protein that Modulates Fibroblast Adhesion and Migration

Meighan-Mantha¹,

Hsu²,

Guo³

et al. 1999

Journal of Biological Chemistry

189

212

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The binding of polypeptide growth factors to their appropriate cell surface transmembrane receptors triggers numerous biochemical responses, including the transcriptional activation of specific genes. We have used a differential display approach to identify fibroblast growth factor-1-inducible genes in murine NIH 3T3 cells. Here, we report that the fibroblast growth factorinducible-14 (Fn14) gene is a growth factor-regulated, immediate-early response gene expressed in a developmental stage-and adult tissue-specific manner in vivo. This gene, located on mouse chromosome 17, is predicted to encode an 129-amino acid type Ia membrane protein with no significant sequence similarity to any known protein. We have used two experimental approaches, direct fluorescence microscopy and immunoprecipitation analysis of biotinylated cell surface proteins, to demonstrate that Fn14 is located on the plasma membrane. To examine the biological consequences of constitutive Fn14 expression, we isolated NIH 3T3 cell lines expressing variable levels of epitope-tagged Fn14 and analyzed their phenotypic properties in vitro. These experiments revealed that Fn14 expression decreased cellular adhesion to the extracellular matrix proteins fibronectin and vitronectin and also reduced serum-stimulated cell growth and migration. These results indicate that Fn14 is a novel plasma membrane-spanning molecule that may play a role in cell-matrix interactions.Complex cellular processes such as proliferation, migration, differentiation, and apoptosis are regulated in part by a diverse group of molecules known as polypeptide growth factors. These factors act by binding and thereby activating specific transmembrane receptor tyrosine kinases. The activation of cell surface receptors by polypeptide ligands triggers downstream intracellular events, including the stimulation of protein phosphorylation cascades and the transcriptional activation of numerous genes (1, 2). Many mitogen-inducible genes have been identified, and they encode a diverse group of proteins including transcription factors, protein kinases and phosphatases, cell cycle regulators, and cytoskeletal and extracellular matrix proteins (2, 3). A recent study using cDNA microarray technology has demonstrated that Ͼ500 genes are transcriptionally activated after serum stimulation of quiescent human fibroblasts and that a subset of these genes encode proteins implicated in the wound healing process in vivo (3).Our laboratory has been studying fibroblast growth factor-1 (FGF-1) 1 -regulated gene expression in murine NIH 3T3 cells. FGF-1 (also referred to as acidic FGF) is one of the most extensively characterized members of the FGF family of heparin-binding proteins (4 -6). It is a potent mitogenic, chemotactic, angiogenic, and neurotrophic factor both in vitro and in vivo. These cellular responses are mediated via high affinity binding to a family of related membrane-spanning tyrosine kinase receptors (4 -6). We have shown by Northern blot hybridization analysis that FGF-1 stimulation of quies...

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Section: Methodsmentioning

confidence: 99%

The Mitogen-inducible Fn14 Gene Encodes a Type I Transmembrane Protein that Modulates Fibroblast Adhesion and Migration

Meighan-Mantha¹,

Hsu²,

Guo³

et al. 1999

Journal of Biological Chemistry

189

212

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“…The transformed non-tumorigenic rat liver cell line NT 27 was used for FGF-1 -stimulation experiments following the method described by Hsu et al [18]. For obtaining a relatively quiescent cell population, confluent cells were either incubated for 120 h in normal medium (control group 1) or for 96 h in medium containing a reduced concentration of serum (0.5 %).…”

Section: Methodsmentioning

confidence: 99%

Further Characterization of a Rat Hepatoma‐Derived Aldose‐Reductase‐Like Protein

Zeindl‐Eberhart

Jungblut

Otto

et al. 1997

European Journal of Biochemistry

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A protein detected in N-methyl-N-nitrosourea-initiated rat hepatomas by two-dimensional electrophoresis at 35 kDdpI 7.4 was identified in a previous study by internal amino acid micro sequencing as an aldose-reductase-like protein [Zeindl-Eberhart, E., Jungblut, P. R., Otto, A. & Rabes, H. M. (1994) Identification of tumor-associated protein variants during rat hepatocarcinogenesis, J. Biol. Chem. 269, 14 589-14 5941. Two-dimensional electrophoresis of rat lens proteins revealed a spot at 37 kDa/pI 6.8 that showed a high degree of identity (98.5%) with rat lens aldose reductase after amino acid sequencing and 80% sequence identity to the rat-hepatoma-derived aldose-reductase-like protein. This suggests that hepatomaderived aldose-reductase-like protein and rat lens aldose reductase are related proteins encoded by different genes. A different expression profile of these proteins was found in various rat organs. Rat lens aldose reductase is present, in addition to in lens, in heart, brain, muscle, lung, duodenum, kidney, spleen and bone marrow, while the hepatoma-derived aldose-reductase-like protein is found preferentially in hepatomas and in embryonic liver. Though different in organ expression, an identical response was found for both proteins after stimulation with fibroblast growth factor-1 and after exposure to increased glucose concentrations. Since rat hepatoma-derived aldose-reductase-like protein is expressed in embryonic, but not in adult liver, it is assumed that it is expressed in hepatomas as a functionally active embryonal type of aldose reductase during hepatocarcinogenesis. Immunohistochemistry revealed that the hepatomaderived aldose-reductase-like protein is expressed already in the preneoplastic stage of hepatocarcinogenesis and might potentially serve as a marker enzyme in early hepatic neoplasia.Keywords: two-dimensional electrophoresis ; rat hepatoma; aldose-reductase-like protein.Misprogramming of genetic information in cancer may lead to quantitative andor qualitative protein alterations. Such changes can be studied by high resolution two-dimensional electrophoresis [ 1-31, in combination with ultrasensitive silver staining [4, 51. Two-dimensional electrophoresis has successfully been used to detect tumor associated proteins in chemically induced rat hepatomas [6-121.We detected previously several protein variants in N-methyl-N-nitrosourea-induced rat hepatomas by two-dimensional electrophoresis [ I l l and identified one of these variants (spot 17) by amino acid sequencing as an enzyme apparently related to rat lens aldose reductase (AR) [13]. In a different system, NIH 3T3 mouse fibroblasts, it was shown that fibroblast growth factor-I (FGF-I) induced temporarily an increased level of FGF-1-regulated (FR-1) mRNA [14]. The amino acid sequence deduced from FR-1 mRNA revealed 97% sequence identity [I41 to the rat hepatoma-derived AR-like protein of chemically induced hepatomas [ 131. These findings of putative relations between rat hepatoma-derived AR-like protein, rat lens AR and FR-1 prot...

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“…(Hsu et al, 1993; Pizer et al, 1997) Based on these observations, it has been hypothesized that de novo lipid synthesis is critical to cell-cycle progression in non-cancer cells such as fibroblasts. (Kuhajda, 2000) However, quantitative assessments of the contribution of de novo lipid synthesis to intracellular palmitate in proliferating non-cancer cells have been limited.…”

Section: Introductionmentioning

confidence: 99%

Exogenous Fatty Acids Are the Preferred Source of Membrane Lipids in Proliferating Fibroblasts

Yao

Fowle-Grider

Mahieu

et al. 2016

Cell Chemical Biology

114

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Summary Cellular proliferation requires formation of additional cellular membrane material, and the current thinking in the field is that lipids required for this new membrane formation are mostly synthesized de novo. Here we measured the contribution of de novo lipid synthesis in proliferating and contact-inhibited fibroblasts and find that proliferating fibroblasts prefer exogenous palmitate over de novo synthesis. We determined that when exogenous palmitate is provided in culture media at physiological concentrations, de novo synthesis accounts for only ~10% of intracellular palmitate in proliferating fibroblasts, as well as HeLa and H460 lung cancer cells. Blocking fatty acid uptake decreased the rate of fibroblast, HeLa, and H460 cell proliferation, while supplementing media with exogenous palmitate resulted in decreased glucose uptake and rendered cells less sensitive to glycolytic inhibition. Thus, our results suggest that cells scavenging exogenous lipids may be less susceptible to both glycolytic and lipogenic inhibitors.

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Fibroblast Growth Factor-1 Induces Phosphofructokinase, Fatty Acid Synthase and Ca2+-ATPase mRNA Expression in NIH 3T3 Cells

Cited by 41 publications

References 0 publications

The Mitogen-inducible Fn14 Gene Encodes a Type I Transmembrane Protein that Modulates Fibroblast Adhesion and Migration

The Mitogen-inducible Fn14 Gene Encodes a Type I Transmembrane Protein that Modulates Fibroblast Adhesion and Migration

Further Characterization of a Rat Hepatoma‐Derived Aldose‐Reductase‐Like Protein

Exogenous Fatty Acids Are the Preferred Source of Membrane Lipids in Proliferating Fibroblasts

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