Demonstration of Basic Fibroblast Growth Factor High and Low Affinity Binding Sites in Human Breast Cancer Cell Lines

Peyrat, Jean‐Philippe; Hondermark, Hubert; Louchez, M M; Boilly, B

doi:10.3727/095535491820873803

Cited by 17 publications

(6 citation statements)

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“…The K, of the high-affinity receptor for bFGF on the myoephitheliallike Rama 711 cells, 13 pM, (Table I), is similar to that observed for the myoepithelial-like cell lines Rama 37-E8 and Rama 29, both of which have been derived from benign mammary tumours (Fernig et ul., 1990). Since the binding of bFGF to the 0.48 X loh low-affinity receptors (K, 6 nM) (Table I) is abolished by the inclusion of 2 pg/ml heparin in the binding medium, this result suggests that the low-affinity receptors are heparan sulfate glycosaminoglycans similar to those found in other mammary myoepithelial-like cells (Fernig et al, 1992) and endothelial cells (Rifkin and Moscatelli, 1989).…”

Section: Discussionsupporting

confidence: 55%

“…3). These higher-molecular-weight forms of bFGF probably represent translation products of bFGF mRNA which have been initiated from thc alternative CUG codons upstream of the AUG codon that initiates translation of 18 kDa bFGF (Rifkin and Moscatelli, 1989). The levels of bFGF mRNA in the Rama 800 cells are comparable to those found in the Rama 711 myoepithelial-like cells, and significantly higher than the level observed in the epithelial Rama 704 cells.…”

Section: Production Of Hepurin-hinding Growth Fuctors By Normal Und supporting

confidence: 54%

“…Moreover, bFGF is produced by the myoepithelial-like mammary cell lines but not by the epithelial cell lines derived from benign tumours (Barraclough et al, 1990). Since bFGF is a potent mitogen and angiogen (Rifkin and Moscatelli, 1989), properties believed to be conducive to the development of malignant tumours, we investigated whether malignant rat mammary epithelial cell lines may have acquired the ability to synthesize hFGF and its rcceptor(s), despite their inability t o dilfcrentiate to the fully differentiated myocpithelial ccll.…”

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confidence: 99%

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Ectopic production of heparin‐binding growth factors and receptors for basic fibroblast growth factor by rat mammary epithelial cell lines derived from malignant metastatic tumours

Fernig

Barraclough

et al. 1993

Intl Journal of Cancer

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A rat mammary (Rama) epithelial cell line, Rama 704, derived from normal rat mammary gland does not possess any detectable cell-surface receptors for basic fibroblast growth factor (bFGF), produces a barely detectable level of bFGF mRNA and does not contain detectable levels of bFGF-like activity. Similar results have been obtained with the Rama 37 epithelial cells derived from benign tumours. However, 4 independently isolated epithelial cell lines derived from malignant rat mammary tumours and their metastases possess receptors for bFGF and contain between 2 ng and 9 ng heparin-binding, growth-stimulatory activity per 10(6) cells. The weakly metastatic Rama 600 cells possess high- and low-affinity receptors for bFGF, (KD 20 pM and 8 nM, respectively), while the moderately metastatic Rama 800 cells possess only high-affinity receptors (KD 40 pM). The moderately metastatic C18PLN and 267LU cells, derived from metastases arising from benign Rama 37 cells which had been transfected with DNA from the malignant Rama 800 cells, also possess only high-affinity receptors (KD 36 pM and 80 pM, respectively). Our results show that within the Rama system there is a correlation between the appearance of heparin-binding growth factors and of high-affinity but not low-affinity receptors for bFGF with the malignant phenotype.

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Section: Discussionsupporting

confidence: 55%

Section: Production Of Hepurin-hinding Growth Fuctors By Normal Und supporting

confidence: 54%

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confidence: 99%

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Ectopic production of heparin‐binding growth factors and receptors for basic fibroblast growth factor by rat mammary epithelial cell lines derived from malignant metastatic tumours

Fernig

Barraclough

et al. 1993

Intl Journal of Cancer

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“…MCF-7 and HBL-100 are hormone-sensitive lines, but BT-20 are a hormone-insensitive one (indications from ATCC references and experiments of two groups from our laboratory, Peyrat et al, 1991;El Yazidi et al, 1998). They were cultured in MEM supplemented with 10% Fetal Calf Serum (FCS), 2 mM glutamine, 100 U-100 mg/ml penicillin-streptomycin, 1% non essential amino acids (all purchased from Eurobio), and 5 mg/ml insulin (Endopancrine, Organon).…”

Section: Cell Culturesmentioning

confidence: 99%

Steroid hormones modulate H19 gene expression in both mammary gland and uterus

et al. 1999

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H19 is an imprinted and developmentally regulated gene whose product remains apparently untranslated. In a previous study on breast adenocarcinomas, we reported that overexpression of the H19 gene was signi®cantly correlated with the presence of steroid receptors, suggesting the putative role of hormones in H19 transcription. To determine the mode of steroid action, we have detected levels of H19 RNA synthesis during mammary gland development by in situ hybridization (ISH): two peaks of H19 transcription occur during puberty and pregnancy. Furthermore, we demonstrated by ISH that in the uterus H19 RNA synthesis is high during estrus and metestrus phases. To test steroid control of H19 transcription, ovariectomized and adrenalectomized mice were supplemented, 1 week after surgery, with 17-b-estradiol (E2, 20 mg/kg/day), progesterone (P, 1 mg/kg/day) or corticosterone (B, 0.3 mg/ kg/day) for 2 weeks. According to ISH data, E2 and to a lesser extent B stimulated H19 transcription in the uterus, whereas P inhibited it. To con®rm the in vivo results, in vitro experiments were performed using cultures of MCF-7 cells (a hormone-sensitive mammary cell line). E2 stimulated the endogenous H19 gene of this cell line and tamoxifen inhibited this e ect. Furthermore, we performed transient cotransfections in MCF-7, in HBL-100 (another hormone-sensitive mammary cell line) and in BT-20 (a hormone-insensitive mammary cell line) with various constructs of ERa (WT or mutated) and PR-A, in presence or absence of steroid hormones. We demonstrated that ERa up-regulated the H19 promoter in MCF-7 and in HBL-100, whereas PR-A did not have any e ect per se. Moreover, in MCF-7, PR-A antagonized clearly the ERa-mediated promoter enhancement, but in HBL-100 this counteracting e ect on the ERa up-regulation was not found. Interestingly, the same experiments performed in BT-20 cell line provided very similar results as those obtained in MCF-7 cells, with a clear down-regulation mediated by PR-A on the H19 promoter. All these in vitro data are in agreement with in vivo results. In addition, data obtained with ERa mutants indicate that H19 promoter activation is both ligand-dependent and ligand-independent. We have thus demonstrated that H19 gene expression is controlled by steroid hormones; furthermore, this gene is highly expressed in hormone-sensitive organs when the hormonal stimulation is accompanied with a morphological repair.

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“…However, these two last breast cell lines possess high affinity membrane receptors for FGF-2 [Peyrat et al, 1991[Peyrat et al, , 1992Lucqmani et al, 1992;Jacquemier et al, 1994;Pennault-Llorca et al, 1995;Johnston et al, 1995]. Low affinity sites for FGF-2 are present in higher concentration on MDA-MB-231 membranes than on MCF-7 cells as shown by a radioreceptor assay [Peyrat et al, 1991[Peyrat et al, , 1992. In the present study, we found that HBL-100 and MDA-MB-231 cells bind higher quantities of FGF2 to their HSPG than MCF-7 cells.…”

Section: Discussionmentioning

confidence: 99%

Production of sulfated proteoglycans by human breast cancer cell lines: Binding to fibroblast growth factor-2

Delehedde¹,

Deudon

Boilly³

et al. 1997

J. Cell. Biochem.

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The cellular distribution and nature of proteoglycans synthesised by human breast cancer cells in culture were studied. Proteoglycans were labelled with [35S] sulfate, purified, and characterised after ion-exchange chromatography followed by gel-filtration chromatography and treatment with glycosaminoglycan degrading enzymes. Proteoglycans were isolated from the culture medium and from cell layers of the hormono-dependent well-differentiated MCF-7 cell line, the hormono-independent poorly-differentiated MDA-MB-231 and the HBL-100 cell line which is derived from non malignant breast epithelium. HBL-100 and MDA-MB-231 cells produced larger amounts of proteoglycans which had a lower degree of sulfation than MCF-7 cells. Gel-filtration chromatography on Sepharose CL-6B indicated that HBL-100 and MDA-MB-231 cells accumulated cell surface heparan sulfate proteoglycans (HSPG), with a high apparent molecular weight (Kav 0.1). In contrast, the MCF-7 cell monolayers synthesised small sulfated macromolecules (Kav 0.4) which possessed mostly chondroitin sulfate chains. Moreover, considerable differences in the nature of the sulfated proteoglycans released into the culture medium of these breast epithelial cell lines were observed. MCF-7 cells released into the culture medium HSPG as the main proteoglycan component while MDA-MB-231 and HBL-100 cells released mainly chondroitin sulfate proteoglycans. In these three cell lines, medium-released sulfated macromolecules have a higher hydrodynamic size than cell-associated ones. Proteoglycans purified by ion-exchange chromatography were tested for their ability to bind 125I FGF-2. We demonstrated that HBL-100 and MDA-MB-231 cells bind more FGF-2 to their heparan sulfate proteoglycans than MCF-7 cells. Taken together, these results suggest that differences in proteoglycan synthesis of human breast epithelial cells could be responsible for differences in their proliferative and/or invasive properties.

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Demonstration of Basic Fibroblast Growth Factor High and Low Affinity Binding Sites in Human Breast Cancer Cell Lines

Cited by 17 publications

References 0 publications

Ectopic production of heparin‐binding growth factors and receptors for basic fibroblast growth factor by rat mammary epithelial cell lines derived from malignant metastatic tumours

Ectopic production of heparin‐binding growth factors and receptors for basic fibroblast growth factor by rat mammary epithelial cell lines derived from malignant metastatic tumours

Steroid hormones modulate H19 gene expression in both mammary gland and uterus

Production of sulfated proteoglycans by human breast cancer cell lines: Binding to fibroblast growth factor-2

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