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PubMed2001

The effects of sialic acid on the gene expression of fibrillar collagens: different changes in normal and fibrotic scar derived fibroblasts.

OBJECTIVE: To investigate the effect of free sialic acid on collagen gene expression in fibroblasts. DESIGN: Cell culture study. SETTING: University hospital, Finland. CELL LINES: Human granulation tissue fibroblasts, human hypertrophic scar fibroblasts and human keloid fibroblasts. INTERVENTIONS: Treatment of cell cultures with 3 microM, 30 microM and 300 microM N-acetyl-neuraminic acid. MAIN OUTCOME MEASURES: The measurement of steady state level of mRNA for type I and type III collagen. RESULTS: Fibroblast lines react dissimilarly under the influence of sialic acid. Granulation tissue fibroblasts showed decrease in the gene expression of type I and III collagen, while keloid fibroblasts contrastingly showed an increase. Hypertrophic scar derived fibroblasts showed no change. CONCLUSIONS: Sialic acids may decrease collagen gene expression in granulation tissue and that disturbed wound healing in diabetics and smokers may in part be due to direct effect of sialic acids on fibroblasts. Sialic acids may in part induce keloid formation.

Cells, Cultured↗

PubMedDate not supplied

Effect of activated human mast cells and mast cell-derived mediators on proliferation, type I collagen production and glycosaminoglycans synthesis by human dermal fibroblasts.

Although an increased number of mast cells in fibrotic tissues such as scleroderma, keloid or healing wound has been highlighted, it is still unclear whether or not mast cells are fibrogenic. The aim of the present study is to determine whether functionally active human mast cells can provide human dermal fibroblasts directly with fibrogenic properties. In order to examine the effects of IgE-mediated mast cell activation on fibroblast proliferation and synthesis of type I collagen, we utilized an in vitro defined system in which cultured human mast cells were co-cultured with human dermal fibroblasts. We also employed a three-dimensional fibroblast culture system using supplementation of L-ascorbic acid as an assay system to investigate the effects of mast cell-derived mediators on synthesis of glycosaminoglycans by human fibroblast. Fibroblast proliferation was actively stimulated with IgE-activated mast cells. However, this stimulatory effect was canceled in co-cultures with a higher number of IgE-activated mast cells. In the presence of a higher number of activated mast cells, the fibroblast cell layer was destroyed, in contrast to an intact cell layer in the presence of same number of the mast cells without activation. Type I collagen synthesis was unchanged in fibroblasts co-cultured with mast cells. The total amount of main disaccharide units, particularly DELTADi-HA, was increased when fibroblasts were exposed to histamine. Thus, we conclude that other factors, in addition to mast cells, are important in the development of human tissue fibrosis or sclerosis.

Cell Division↗

PubMed2002

Rat periodontal fibroblast responses to bacterial lipopolysaccharide in vitro.

Rat periodontal ligament fibroblasts and gingival fibroblasts were stimulated with lipopolysaccharide from Escherichia coli. Cell proliferation and both interleukin-1beta and nitric oxide production were determined by a colorimetric, enzyme-linked immunosorbent assay and the Griess reaction, respectively. The results showed that at lipopolysaccharide concentration up to 100 ng per well, the cell proliferation of periodontal ligament fibroblasts was higher than that of gingival fibroblasts. No significant difference between the levels of interleukin-1beta produced by periodontal ligament and gingival fibroblasts can be observed. At high concentration of lipopolysaccharide (1000 ng/well), the levels of nitric oxide in the periodontal ligament fibroblasts cultures were higher than those in the gingival fibroblasts cultures. These results suggest those rat periodontal ligament and gingival fibroblasts may respond differently to lipopolysaccharide and, thus, may be different periodontal fibroblast subpopulations.

Animals↗

PubMed2002

Partial restoration of the keratocyte phenotype to bovine keratocytes made fibroblastic by serum.

PURPOSE: To determine whether keratocytes made fibroblastic in vitro by addition of fetal bovine serum to the medium regain the keratocyte phenotype after culture in serum-free medium. METHODS: Collagenase-isolated keratocytes from bovine corneas were plated in DMEM/F-12 containing 1% horse plasma, to allow cell attachment, and then cultured until day 4 in either DMEM/F-12 alone, to retain the keratocyte phenotype, or in DMEM containing 10% fetal bovine serum, to cause the keratocytes to become fibroblastic. Medium for the fibroblastic cells was replaced on day 4 with serum-free medium, and cells were cultured until day 12. Cell phenotypes were determined on days 4 to 5 and 11 to 12 of culture as follows: (1) by the morphologic appearance on phase-contrast microscopy; (2) by the levels of aldehyde dehydrogenase in the cells, determined by SDS-PAGE and Coomassie blue staining; (3) by the relative synthesis of collagen types I and V, determined by (14)C-proline radiolabeling; (4) by pepsin digestion and analysis of collagen types by SDS-PAGE autoradiography; (5) by relative synthesis of cornea-specific proteoglycan core proteins determined by analysis of chondroitinase- or endo-beta-galactosidase-generated radiolabeled core proteins by SDS-PAGE autoradiography; and (6) by the relative synthesis of keratan sulfate and chondroitin sulfate determined by (35)SO(4) radiolabeling and measuring the sensitivity to endo-beta-galactosidase and chondroitinase ABC. RESULTS: Keratocytes cultured in serum-free medium appeared dendritic and became fibroblastic in appearance when exposed to medium containing serum. Keratocytes and fibroblasts synthesized a similar proportion of collagen types I and V. However, compared with the keratocytes, the fibroblasts possessed no aldehyde dehydrogenase and synthesized significantly higher levels of decorin and significantly lower levels of prostaglandin D synthase (PGDS) and keratan sulfate. Subsequent culture of the fibroblasts in serum-free medium did not restore aldehyde dehydrogenase to keratocyte levels but did restore the cell morphology to a more dendritic appearance and returned the synthesis of decorin, PGDS, and keratan sulfate to keratocyte levels. CONCLUSIONS: The results of these studies indicate that primary cultures of keratocytes made fibroblastic by exposure to serum can return to their keratocyte phenotype in synthesizing extracellular matrix. These results also indicate that the differences in the organization of the collagenous matrix produced by keratocytes and fibroblasts may be related more to the different proteoglycan types than to the collagen types produced.

Aldehyde Dehydrogenase↗

PubMed2000

[Expression of platelet derived growth factor receptor-beta in fibroblasts of keloid].

OBJECTIVE: To investigate the expression and distribution of platelet derived growth factor receptor-beta(PDGFR-beta) in normal skin and keloid and to discuss its biological function in keloid formation. METHODS: 1. To detect the expression and distribution of PDGFR-beta in normal skin and keloid tissue by immunohistochemistry; 2. To detect the receptor expression in vitro by Flow cytometry (FCM); 3. To detect the subcellular distribution of receptor by Laser confocal microscope. RESULTS: 1. Immunohistochemistry showed that normal skin and keloid tissue were almost the same in expression but different in distribution of PDGFR-beta; 2. There was more expression of PDGFR-beta in normal fibroblasts than that in keloid fibroblasts in vitro by FCM; 3. Laser confocal microscope revealed that the PDGFR-beta concentrated on the surface of cell membrane in keloid fibroblasts, but in normal skin fibroblasts, the receptors were coagulated on the nuclear membrane and intranucleus. CONCLUSION: Compared with the fibroblasts in vivo, there was a difference of the PDGFR-beta expression in fibroblasts in vitro, more expression of PDGFR-beta in normal fibroblast than that in keloid fibroblast in vitro; and the subcellular distribution of PDGFR-beta was different in normal skin and keloid fibroblasts. The characteristics of the expression and distribution of PDGFR-beta in keloid may contribute to the formation of keloid.

Cells, Cultured↗

PubMed2002

[Culture of human fibroblasts transfected by human telomerase reverse transcriptase eucaryotic expression plasmid pGRN145 and their biological characteristics in vitro].

OBJECTIVE: To analysis the biological characteristics of human fibroblasts transfected by human telomerase reverse transcriptase (hTERT) eucaryotic expression plasmid pGRN145. METHODS: Fibroblasts from children's foreskin were isolated and cultured in vitro, and the fibroblasts were transfected by pGRN145 with Lipofec-tAMINE PLUS Reagent. After strict screening of hygromycin B, the positive clones were subcultured. The telomerase activity was detected by RT-PCR and TRAP-PCR technique. The cell generation cycle and apoptosis rate were detected by flow cytometry to investigate the proliferative characteristics after transfection, and the chromosome karyotype of transformed cells was analyzed. The collagen secreted by transformed cells was detected by immunohistochemical staining. RESULTS: The morphological properties of fibroblasts did not change obviously after transfection. There were telomerase activity in transfected fibroblasts, while it could not be detected in pre-transfection fibroblasts. The cell generation cycle had no obvious changes between pre-transfection and post-transfection. However, the apoptosis rate of transfected fibroblasts were decreased compared with that of pre-transfection. The fibroblasts transfected by pGRN145 maintained the normal diploid karyotype, as well as the cells could normally secret type I and III collagen. CONCLUSION: The human fibroblasts transfected by pGRN145 has telomerase activity with prolonged life span of culture, which preliminarily proves the availability of establishing standard seeding cell lines of tissue engineering by hTERT plasmid transfection techniques.

Apoptosis↗

PubMed2002

[The distributing characteristics and proliferating activity of fibroblasts from the surrounding skin of keloids].

OBJECTIVE: To investigate whether there are abnormal fibroblasts derived from the surrounding skin of keloids so that a more accurate therapy for keloids could be obtained. METHODS: Samples were taken for cell culture. When primary cells fully covered the culture bottle, the shape and distributing characteristics of fibroblasts were observed under the light microscope. 6-8 passage fibroblasts were selected for comparing the proliferating activity by MTT contrasting color method. RESULTS: The fibroblasts have the same shape in all groups. But the fibroblasts derived from the surrounding skin grow crossly and overlapped just as the fibroblasts from keloids. The proliferating activity of the fibroblasts from surrounding skin is not as high as that from the border of keloids, but is higher than the normal skin fibroblasts derived either from a normal person or a patient with keloid. CONCLUSION: It is likely that there are abnormal fibroblasts in the surrounding skin of keloids.

Adolescent↗

PubMed2003

Characterization and functionality of CXCR4 chemokine receptor and SDF-1 in human corneal fibroblasts.

PURPOSE: The aim of this study was to investigate whether cultured human corneal fibroblasts express functional chemokine CXCR4 receptors on their cell surface and to determine the presence of its specific ligand, SDF-1 (CXCL12), in human corneal fibroblasts. METHODS: Human corneal fibroblast cultures were obtained using human donor corneas. CXCR4 receptors were characterized using binding studies and autoradiography with [125I]SDF-1. The functionality of CXCR4 receptors was assessed by intracellular calcium measurement using a dynamic imaging microscopy system. CXCR4 and SDF-1 mRNA were detected in human corneal fibroblasts using reverse transcriptase polymerase chain reaction (RT-PCR). The CXCR4 protein was detected by western blot analysis. RESULTS: [125I]SDF-1 specifically bound to cultured corneal fibroblasts with a KD value of 8.3+/-1.2 nM. The presence of CXCR4 was confirmed by autoradiography of the radioligand on slices of corneal stroma. SDF-1 induced a rapid and transient intracellular calcium increase in cultured corneal fibroblasts that was blocked by the specific antagonist bicyclam. Moreover, a 48 kDa protein was detected by western blot analysis of corneal fibroblast extracts, using a specific CXCR4 polyclonal antibody. RT-PCR showed the expression of both CXCR4 and SDF-1 mRNAs in human corneal fibroblasts. CONCLUSIONS: These results indicate for the first time that cultured human corneal fibroblasts express the chemokine receptors CXCR4 and its ligand SDF-1. This latter might exert physiological effects on the cornea and could be involved in pathological conditions such as corneal angiogenesis.

Autoradiography↗

PubMed1992

Transfection of SV40-transformed ataxia-telangiectasia fibroblasts with mouse DNA corrects hypersensitivity to neocarzinostatin and activates fibronectin gene expression.

SV40-transformed ataxia-telangiectasia (SV40-AT) fibroblasts were cotransfected with a plasmid carrying the neomycin-resistance gene as well as DNA from primary mouse embryo fibroblasts. The transfected fibroblasts were seeded under selective conditions and neomycin-resistant (neor) colonies were obtained and tested for the effect of the carcinogen neocarzinostatin (NCS) on DNA synthesis. Whereas the primary A-T and SV40-transformed A-T fibroblasts did not respond to carcinogen treatment and continued to synthesize DNA on damaged templates, normal fibroblasts stopped DNA synthesis after NCS treatment. Among the neomycin-resistant colonies, cells of two colonies responded to NCS treatment by the cessation of DNA synthesis like normal fibroblasts. When DNA from such a colony was transfected into SV40-AT cells, four neor colonies were isolated of which one had regained the normal phenotype. This study provides the first clue that mouse DNA can partly correct the A-T genetic defect expressed in SV40-transformed fibroblasts. Two of the neor colonies with the corrected phenotype expressed a 3.5 kb fibronectin RNA that was detectable by a rat fibronectin DNA probe but not by the human fibronectin DNA probe containing the cell attachment sequence. The latter probe did not detect fibronectin mRNA in the SV40-AT cells but detected expression of the 8.6 kb fibronectin RNA in the two neor colonies of transfected SV40-AT fibroblasts in which the response to NCS was repressed. The results suggest that "correction" of the A-T gene defect in SV40-AT fibroblasts might be associated with regulation of human fibronectin gene(s) expression.

Animals↗

PubMed1992

Necessity of extracellular domain of W (c-kit) receptors for attachment of murine cultured mast cells to fibroblasts.

The receptor encoded by the W (c-kit) locus (W receptor) is expressed on the surface of cultured mast cells (CMC) derived from normal (+/+) mice, whereas its ligand encoded by the Sl locus (Sl ligand) is expressed on the surface of fibroblast cell lines derived from murine embryos. Involvement of W receptors and Sl ligands in attachment of CMC to fibroblasts was investigated. CMC were cocultured with fibroblasts; nonattaching CMC were removed and the remaining CMC were counted. CMC derived from mice of the W/W genotype did not express the extracellular domain of W receptors, and attachment of W/W CMC to +/+ fibroblasts was significantly impaired. Fibroblasts derived from embryos of the Sl/Sl genotype did not express Sl ligands, and the attachment of +/+ CMC to Sl/Sl fibroblasts was also impaired. The Wv and W42 alleles are point mutations at the intracellular tyrosine kinase domain. Attachment of either Wv/Wv, W/Wv, or W/W42 CMC to +/+ fibroblasts was comparable with that of +/+ CMC. Moreover, the addition of monoclonal antibody against the extracellular domain of W receptors inhibited the attachment of +/+ CMC to +/+ fibroblasts. Thus, the extracellular domain of W receptors appeared to be necessary for attachment of CMC to fibroblasts.

Alleles↗

PubMed2003

[The inhibition effect of interleukin-1 receptor antagonist on interleukin-1 alpha-induced intercellular adhesion molecule-1 expression on orbital fibroblasts and adhesion of peripheral blood mononuclear cells].

OBJECTIVE: The aim was to identify whether interleukin-1 receptor antagonist (IL-1ra) inhibits interleukin-1 alpha (IL-1 alpha)-induced intercellular adhesion molecule-1 (ICAM-1) expression on cultured orbital fibroblasts, and adhesion of peripheral blood mononuclear cells (PBMC) to orbital fibroblasts, and to investigate the clinical application potential of IL-1ra in the treatment of Graves' ophthalmopathy (GO). METHODS: Cultured orbital fibroblasts from patients with GO and controls were challenged with IL-1 alpha or/and IL-1ra. Immunocytochemical staining was used to examine the changes of ICAM-1 in response to IL-1ra treatment; fluorescent photomicroscope was used to measure the adhesion between the labeled PBMC and orbital fibroblasts. Neutralizing antibody against ICAM-1 was used to demonstrate the role of ICAM-1 in the IL-1 alpha-induced adhesion. RESULTS: IL-1ra inhibits IL-1 alpha-induced ICAM-1 expression in cultured orbital fibroblasts both from GO patients and controls; IL-1ra inhibits IL-1 alpha-induced adhesion of PBMC to orbital fibroblasts in a concentration and time dependent manner. Moreover, a monoclonal anti-human ICAM-1 antibody produced a concentration dependent inhibition of the IL-1 alpha-induced adhesion of PBMC to the fibroblasts. CONCLUSIONS: IL-1ra inhibits IL-1 alpha-induced ICAM-1 expression in cultured orbital fibroblasts and the adhesion of PBMC to fibroblasts. IL-1 alpha-induced ICAM-1 expression may play an important role in the adhesion process. IL-1ra may be useful in the prevention or treatment of GO.

Cell Adhesion↗

PubMed2004

[Accelerated angiogenesis in artificial dermis with allogenic cultured dermal fibroblasts transplantation].

OBJECTIVE: To evaluate the effects of cryopreserved cultured allogenic dermal fibroblasts on angiogenesis and fibroplasia while artificial dermis grafting by spraying the cells on the graft bed. METHODS: Full thickness skin defect was made on the back of Wistar rat, fibroblasts mixed into fibrin glue (fibroblast group) and same amount fibrin glue (control group) were sprayed separately between the wound bed and artificial dermis in cell density of 1.0 x 10(5) cells/cm2 before the artificial dermis was grafted. On day 5 after grafting, the graft and surrounding tissue were examined histologically for angiogenesis and fibroplasia in the dermis and wound bed with hematoxylin eosin stain, VEGF antibody stain, Masson's trichrome stain and India ink stain. Evans blue perfusion method was also used for detecting the angiogenesis quantitatively. RESULTS: In the fibroblast group, the angiogenesis of graft bed was significantly accelerated on day 5 after grafting; the numbers of the newly formed capillaries were 9.64 +/- 2.36/HP in the fibroblast group and 3.88 +/- 1.62/HP in the control group (P < 0.05). And on day 10 after grafting the angiogenesis was accelerated not only in graft bed but also in the artificial dermis when compared with control group, the newly formed capillaries network was clearly observed in the artificial dermis. Otherwise, the synthesis of collagen was increased in the dermis on day 10 after grafting in the fibroblast group when compared with control group. The immunoreactivity of VEGF antibody in the fibroblast group also showed a stronger expression than that in control group on day 5 after grafting, the numbers of positive cells were 46.04 +/- 8.90/HP in the fibroblast group and 30.08 +/- 7.76/HP in the control group(P < 0.05). CONCLUSION: Transplantation of cryopreserved dermal fibroblasts while artificial dermis grafting can accelerate the angiogenesis and fibroplasia in the artificial dermis and graft bed, thereby accelerate the formation of dermal-like tissue in the artificial dermis.

Animals↗

PubMed2004

[Effects of colchicine on synthesis and excretion of cytokines and extracellular matrix by human renal fibroblasts].

OBJECTIVE: Renal interstitial fibrosis is the final common pathway leading to end-stage renal failure for progressive renal disease of various types. The present study was undertaken to add to the knowledge on colchicine's antiinflammatory and antifibrotic properties confirmed by both human and experimental studies. As the main effector cells, fibroblasts have a central role in the pathogenesis of renal fibrosis. This study aimed to investigate the effects of colchicine on the synthesis and excretion of cytokines transforming growth factor-beta1 (TGF-beta1), interleukin-1beta (IL-1beta) and extracellular matrix (collagen III, collagen IV) in human renal fibroblast. METHODS: Various concentrations of colchicine (5.0 micromol/L, 10.0 micromol/L, 20.0 micromol/L, 40.0 micromol/L) were used to pretreat human embryo renal fibroblasts for 1 hour which were cultured in vitro and then stimulated by 10.0 microg/ml of lipopolysaccharide (LPS). After 18 hours, these fibroblasts and their supernatant were collected. The expression of TGF-beta1 mRNA, IL-1beta mRNA in the cells was studied by using RT-PCR, and the excretion of TGF-beta1, IL-1beta, collagen III and collagen IV by the fibroblasts was assessed by ELISA respectively. RESULTS: (1) By pure stimulation with 10.0 micro g/ml LPS, the expression of TGF-beta1 mRNA and IL-1beta mRNA of fibroblasts was up-regulated 3 times (66.1 +/- 1.6 vs. 22.3 +/- 2.0, q = 590.5, P = 0.002) and 4.7 times (22.0 +/- 2.2 vs. 4.7 +/- 0.8, q = 106.8, P = 0.009), respectively. The protein excretion of TGF-beta1 and IL-1beta was remarkably increased as well compared with the control group [TGF-beta1: (516 +/- 14) pg/ml vs. (420 +/- 5) pg/ml (q = 80.3, P = 0.012), IL-1beta: (3.4 +/- 0.3) pg/ml vs. (0.3 +/- 0.1) pg/ml (q = 297.9, P = 0.003)]. (2) Colchicine could inhibit the expression of TGF-beta1 mRNA and protein in a dose-dependent manner. IL-1beta mRNA and protein were both up-regulated by colchicine. (3) LPS could not stimulate the excretion of extracellular matrix by fibroblasts, but the excretion of collagen III and collagen IV was down regulated by colchicine in a dose-dependent manner. CONCLUSION: (1) The expression of TGF-beta1 mRNA and the excretion of TGF-beta1 protein in the fibroblasts was significantly suppressed by colchicine, while the expression of IL-1beta mRNA and the excretion of IL-1beta protein were enhanced. (2) Colchicine has significant inhibitory effect on the excretion of extracellular matrix such as collagen III and collagen IV in fibroblasts.

Colchicine↗

PubMed2004

Autocrine activation by interleukin 1alpha induces the fibrogenic phenotype of systemic sclerosis fibroblasts.

OBJECTIVE: To explore the cellular localization of interleukin 1alpha (IL-1alpha) in cultured fibroblasts from lesional skin of patients with systemic sclerosis (SSc) and to study the role of intracellular IL-1alpha in the activation of fibroblasts. METHODS: Dermal fibroblasts were derived from 12 patients with SSc. Expression of IL-1alpha mRNA was examined using reverse transcriptase-polymerase chain reaction (RT-PCR). The cellular distribution of IL-1alpha was examined by subcellular fractionation, flow cytometry, and immunocytochemistry. A full-length IL-1alpha cDNA was subcloned into the pcDNA3 vector to create sense and antisense-encoding constructs. Normal and SSc fibroblasts were stably transfected with the sense and antisense-encoding constructs, respectively. Stably transfected fibroblast clones were analyzed for the production of procollagen and IL-6 protein by ELISA, alpha1(I) procollagen mRNA by Northern blot hybridization, and proliferation by [3H]thymidine incorporation. RESULTS: SSc-affected fibroblasts constitutively expressed intracellular IL-1alpha, which was predominantly located in the nucleus. Inhibition of IL-1alpha expression in SSc-affected fibroblasts using antisense constructs resulted in decreased proliferation, IL-6 production, and procollagen synthesis. Conversely, overexpression of IL-1alpha in normal fibroblasts resulted in development of the SSc fibroblast phenotype. CONCLUSION: IL-1alpha is an important autocrine fibrogenic factor in SSc, suggesting that inhibition of intracellular IL-1alpha may be a novel strategy for the treatment of SSc.

Autocrine Communication↗

PubMed2005

[Pathological genomics of keloid fibroblastic cells].

OBJECTIVE: Keloids result from the abnormal repair of the tissues after skin injuries where the pathological overgrowth of large and active fibroblastic cells expands beyond the boundaries of the initiating wound. Imbalanced expression of genes with an as yet unknown regulatory mechanism seems to result in the hypertrophic development of fibroblastic cells and over-productions of collagen. To get information as to genes which function in the actively growing keloid fibroblasts, we have applied a gene expression DNA-microarray technique by analyzing broad range of genes at once in a systematic fashion. METHODS: Differential gene expressions of keloid fibroblastic cell lines against a normal skin fibroblastic cell line, all of the cell lines had been propagated in our lab, were analyzed using a cDNA-microarray technique. mRNA was extracted from the control normal skin cells and the two lines of keloid fibroblastic cells, one from ear-lobe keloid tissue and the other from chest keloid tissue, was subjected to a DNA microarray analysis which includes 1 100 human genes (TaKaRa Intelli Gene Human CHIP 1K Set I) . RESULTS: 8 genes were found to be expressed exclusively in ear-lobe keloid fibroblastic cell lines. Cells from chest keloid were detected to express 17 genes, specifically. Coagulation factor II (thrombin) receptor gene, KIAA0367 protein gene, and matrilin-2 gene were found to be the most commonly expressed genes in the keloid cells. Suppressor genes, like melanoma differentiation associated gene-7, Mda-7 (U16261), were expressed in normal skin fibroblasts but were not expressed in keloid fibroblasts may be implicated in the pathogenesis of the keloid lesions. CONCLUSIONS: Genes expressed specifically in keloid cells may be an adequate pathological diagnostic marker for keloids. Further, Identification of genes that cause cells to develop keloid lesions leads us to gene therapy and prevention of keloids.

Adult↗

PubMed1991

[Interaction of lung cancer cells and fibroblasts].

Fibroblasts are found to secrete several factors, but complex interactions between cancer cells and fibroblasts are not well known. In this report, it was discussed how fibroblasts derived from lung cancer tissues affected lung cancer cells in vitro and in vivo. As results, the growth of 4 of eight cancer cells was promoted by any medium conditioned by fibroblasts (C.M) in vitro. Fifty percent of the 12 fibroblasts had some effect on promoting the growth of cancer cells, but had no effect on suppressing cancer cells. The C.Ms promoting the growth of cancer cells in vitro exerted the same effect in nude mice, too. Coinoculation of fibroblasts and cancer cells into nude mice gained higher tumorigenicity than cancer cells alone with no relation to their subclasses. It was suggested that to examine the effect on promoting the growth of cancer cells by fibroblasts derived from lung cancer tissues in vitro was able to prospect how fibroblasts affected cancer cells in vivo and fibroblasts made some good condition to transplanted cancer cells into nude mice by means different from growth factors.

Animals↗

PubMed1991

Fibronectin release by systemic sclerosis and normal dermal fibroblasts in response to TGF-beta.

To determine whether enhanced matrix synthesis by systemic sclerosis (SSc) fibroblasts in vitro is due to increased responsiveness to transforming growth factor-beta (TGF-beta), fibronectin release by SSc and normal fibroblasts (7 pairs) was measured at various concentrations of TGF-beta. In the absence of TGF-beta, SSc fibroblasts released 30 +/- 22% more fibronectin than normal fibroblasts. While both SSc and normal fibroblasts increased fibronectin release at all concentrations of TGF-beta tested, the percentage increases were not statistically greater for the SSc fibroblasts even though 4 of the SSc fibroblasts strains were selectively sensitive to low concentrations of TGF-beta. TGF-beta increased cell numbers of both SSc and normal strains equally. Our data confirm abnormal regulation of fibronectin gene expression in SSc fibroblasts and suggest increased sensitivity to TGF-beta by some SSc fibroblast strains.

Blotting, Western↗

PubMed1991

Transforming growth factor-beta regulation of IL-6 production by unstimulated and IL-1-stimulated human fibroblasts.

We characterized the ability of transforming growth factor-beta 1 (TGF-beta 1) and transforming growth factor-beta 2 (TGF-beta 2) to regulate IL-6 production by unstimulated and rIL-1-stimulated lung fibroblasts. rTGF-beta 1-, purified TGF-beta 1-, and purified TGF-beta 2-stimulated fibroblasts produced IL-6 bioactivity as assessed with the B9 hybridoma proliferation assay. These TGF-beta moieties also bidirectionally regulated the IL-6 production of rIL-1-stimulated fibroblasts. The addition of TGF-beta to cultures in which fibroblasts were vigorously stimulated with rIL-1 resulted in an inhibition of fibroblast IL-6 production and mRNA accumulation. In contrast, the addition of TGF-beta to cultures in which fibroblasts were incubated with suboptimal concentrations of rIL-1 resulted in a synergistic increase in IL-6 production and mRNA accumulation [corrected]. Nuclear run-on analysis demonstrated that IL-6 gene [corrected] transcription was synergistically augmented when rTGF-beta 1 was combined with suboptimal concentrations of rIL-1. These studies demonstrate that TGF-beta stimulates fibroblast IL-6 production. They also show that TGF-beta can augment or inhibit the IL-6 production of IL-1-stimulated fibroblasts. Lastly, [corrected] they demonstrate that the stimulatory effects of TGF-beta are, at least partially, mediated by alterations in IL-6 gene transcription. TGF-beta may be an important regulator of IL-6 production. stimulated fibroblasts. Last, they demonstrate that the stimulatory effects of TGF-beta are, at least partially, mediated by alterations in IL-6 gene transcription. TGF-beta may be an important regulator of IL-6 production.

Cells, Cultured↗