E1A stimulates FGF-2 release promoting differentiation of primary endothelial cells

Giampietri, Claudia; Levrero, Massimo; Felici, Angelina; D’Alessio, Alessio; Capogrossi, M. C.; Gaetano, Carlo

doi:10.1038/sj.cdd.4400654

Cited by 9 publications

(5 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Primary bovine aorta endothelial cells (BAECs) were obtained as reported previously 42 and were cultured at 37°C in a 5% CO 2 atmosphere in Dulbecco modified Eagle medium (DMEM; Hyclone, Logan, UT) supplemented with 2 mM L-glutamine, 100 IU/mL penicillin-streptomycin (Gibco, Gaithersburg, MD), and 10% fetal calf serum (FCS; Hyclone). BAEC preparation purity was evaluated by low-density lipoprotein uptake and was consistently greater than 97%.…”

Section: Cell Culturementioning

confidence: 99%

“…42 Briefly, 200 L reconstituted extracellular matrix protein (Matrigel), growth factor-reduced and without phenol red (Collaborative Research, Bedford MA), was applied to 24-well culture plates and was incubated at 37°C for 60 minutes. BAEC (8 ϫ 10 4 in 1 mL medium) were seeded on Matrigel-coated wells in the presence of 0.1% BSA, 1% FCS, or bFGF (10 ng/mL), or bFGF/PDGF-BB (10 ng/mL each), or PDGF-BB alone (10 ng/mL), and were kept at 37°C.…”

Section: Differentiation Assaymentioning

confidence: 99%

See 1 more Smart Citation

Platelet-derived growth factor inhibits basic fibroblast growth factor angiogenic properties in vitro and in vivo through its α receptor

Marchis

Ribatti

Giampietri

et al. 2002

Blood

View full text Add to dashboard Cite

Basic fibroblast growth factor (bFGF) and platelet-derived growth factor-BB (PDGF-BB) modulate vascular wall cell function in vitro and angiogenesis in vivo. The aim of the current study was to determine how bovine aorta endothelial cells (BAECs) respond to the simultaneous exposure to PDGF-BB and bFGF. It was found that bFGF-dependent BAEC migration, proliferation, and differentiation into tubelike structures on reconstituted extracellular matrix (Matrigel) were inhibited by PDGF-BB. The role played by PDGF receptor ␣ (PDGF-R␣) was investigated by selective stimulation with PDGF-AA, by blocking PDGF-BB-binding to PDGF-R␣ with neomycin, or by transfecting cells with dominant-negative forms of the receptors to selectively impair either PDGF-R␣ or PDGF-R␤ function. In all cases, PDGF-R␣ impairment abolished the inhibitory effect of PDGF-BB on bFGF-directed BAEC migration. In addition, PDGF-R␣ phosphorylation was increased in the presence of bFGF and PDGF, as compared to PDGF alone, whereas mitogen-activated protein kinase phosphorylation was decreased in the presence of PDGF-BB and bFGF compared with bFGF alone. In vivo experiments showed that PDGF-BB and PDGF-AA inhibited bFGF-induced angiogenesis in vivo in the chick embryo chorioallantoic membrane assay and that PDGF-BB inhibited bFGF-induced angiogenesis in Matrigel plugs injected subcutaneously in CD1 mice. Taken IntroductionThe endothelial layer represents a physical and chemical barrier between the vessel lumen and the underlying tissues. Endothelial cells (ECs) exert a variety of functions and modulate underlying smooth muscle cells by releasing molecules with vasoactive and growth-regulatory properties. 1 Endothelial cells present an active replication phenotype in vitro, but in vivo they are quiescent. 2 The different expression of membrane-bound receptors 3,4 and the in vivo action of specific inhibitors 5-7 may account, at least in part, for the different replication pattern observed.Basic fibroblast growth factor (bFGF) is a potent EC growth factor; it is known to induce a proangiogenic phenotype in ECs and is released under acidosis conditions that induce EC protection from apoptosis. 8 bFGF plays a critical role in physiologic and pathologic angiogenesis, including tumor angiogenesis. 9,10 It exerts its functions by direct action, 11 by inducing vascular endothelial growth factor (VEGF) synthesis, 12 or by potentiating VEGF activity. 13,14 Platelet-derived growth factor (PDGF) is a growth factor known to be active on ECs. Three PDGF isoforms have been identified as disulfide-linked dimers, namely PDGF-AA, PDGF-BB, and PDGF-AB, expressed by ECs under various conditions. 15-19 They interact with different affinity with 2 tyrosine-kinase receptors, PDGF-R␣ and PDGF-R␤, which are expressed on ECs in normal 4,20-22 and in pathologic conditions. [23][24][25][26] Recently, PDGF-C and PDGF-D isoforms were also identified. 27,28 PDGF isoforms are reported to exert mitogenic and chemotactic action on EC, although PDGF-AA appears to be less potent or inactive...

show abstract

Section: Cell Culturementioning

confidence: 99%

Section: Differentiation Assaymentioning

confidence: 99%

Platelet-derived growth factor inhibits basic fibroblast growth factor angiogenic properties in vitro and in vivo through its α receptor

Marchis

Ribatti

Giampietri

et al. 2002

Blood

View full text Add to dashboard Cite

show abstract

“…BAEC were derived from adult bovine aorta and cultured as previously reported (40). Cell purity was evaluated by DiI-Ac-LDL uptake and was consistently Ͼ97%.…”

Section: Cell Isolation and Cell Culturementioning

confidence: 99%

Identification of a Novel Domain of Fibroblast Growth Factor 2 Controlling Its Angiogenic Properties

Facchiano

Russo

Facchiano

et al. 2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

Fibroblast growth factor 2 (FGF-2) is a potent factor modulating the activity of many cell types. Its dimerization and binding to high affinity receptors are considered to be necessary steps to induce FGF receptor phosphorylation and signaling activation. A structural analysis was carried out and a region encompassing residues 48 -58 of human FGF-2 was identified, as potentially involved in FGF-2 dimerization. A peptide (FREG-48 -58) derived from this region strongly and specifically inhibited FGF-2 induced proliferation and migration of primary bovine aorta endothelial cells (BAEC) in vitro, and markedly reduced FGF-2-dependent angiogenesis in two distinct in vivo assays. To further investigate the role of region 48 -58, a polyclonal antibody raised against FREG-(48 -58) was tested and was found to block FGF-2 action in vitro. Human FGF-2 has three histidine residues, one falling within the region 48 -58. Chemical modification of histidine residues blocked FGF-2 activity and FREG-(48 -58) inhibitory effect in vitro, indicating that histidine residues, in particular the one within FREG-(48 -58) region, play a crucial role in the observed activity. Additional experiments showed that FREG-(48 -58) specifically interacted with FGF-2, impaired FGF-2-interaction with itself, with heparin and with FGF receptor 1, and inhibited FGF-2-induced receptor phosphorylation and FGF-2 internalization. These data indicate for the first time that region 48 -58 of FGF-2 is a functional domain controlling FGF-2 activity.

show abstract

“…54 Briefly, total RNA was extracted by using TRIZOL reagent (Life Technologies, Milano, Italy) according to the manufacturer's instructions. RNA was subjected to electrophoresis through a 1% agarose gel containing formaldehyde and was transferred onto a nylon membrane (Schleicher & Schuell, Keene NH, USA).…”

Section: Northern Blottingmentioning

confidence: 99%

FLIP is expressed in mouse testis and protects germ cells from apoptosis

et al. 2003

Self Cite

View full text Add to dashboard Cite

Apoptosis control in adult testis is crucial to achieve normal spermatogenesis. In this study c-FLIP, an apoptosis-modulating protein, was investigated. In Western blot and immunohistochemical analyses, the 55 KDa c-FLIP long isoform (c-FLIP L ) was found to be expressed strongly in spermatocytes and spermatids, at low levels in spermatogonia and at almost undetectable levels in Sertoli cells. This expression pattern was confirmed by Northern blot analyses. Further experiments carried out on GC-1spg germ cell line revealed that reducing c-FLIP L expression increases Fas-dependent apoptosis. Conversely, restoring c-FLIP L expression reduces this response to control levels. Caspase-10 expression was found to match c-FLIP L expression pattern; further, caspase-10 activation upon anti-Fas treatment inversely correlated with c-FLIP L expression. Finally, TUNEL staining of seminiferous tubules incubated with anti-Fas antibody showed that apoptosis occurs mostly in basally located germ cells, indicating that such cells, expressing low levels of c-FLIP L , are sensitive to Fas-mediated apoptosis. These data indicate for the first time that c-FLIP L might control germ cell apoptosis and caspase activity in the adult testis.

show abstract

E1A stimulates FGF-2 release promoting differentiation of primary endothelial cells

Cited by 9 publications

References 61 publications

Platelet-derived growth factor inhibits basic fibroblast growth factor angiogenic properties in vitro and in vivo through its α receptor

Platelet-derived growth factor inhibits basic fibroblast growth factor angiogenic properties in vitro and in vivo through its α receptor

Identification of a Novel Domain of Fibroblast Growth Factor 2 Controlling Its Angiogenic Properties

FLIP is expressed in mouse testis and protects germ cells from apoptosis

Contact Info

Product

Resources

About