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B Westermark

Publications and source records attributed to B Westermark.

At least 73 records · Page 4Linked to original sources
PubMed1991

Epidermal growth factor stimulates thyroid follicle neogenesis in collagen gel culture.

The mechanism involved in the neoformation of thyroid follicles is poorly understood. In the present study, whole porcine thyroid follicles were cultured as "miniorgans", embedded within collagen gels. Incubation was performed up to 4 days with or without EGF (10 ng/ml) and TSH (2 mU/ml). A single dose of 3H-thymidine was added at the start of the experiments in some cases. Light microscopy and autoradiography was performed. EGF induced a dramatic migration of follicles cells; these were seen to back out from the mother follicle and formed microfollicles with normal polarity including microvilli at the apical border. As evident from the analysis of 3H-thymidine incorporation, most microfollicles were comprised of newly divided cells. The results infer a new mechanism for the neoformation of follicles in the thyroid.

Animals↗
PubMed1990

Expression of messenger RNAs for platelet-derived growth factor and its receptors in human sarcoma cell lines.

Growth factors of the platelet-derived growth factor (PDGF) family have been thought to possess autocrine functions in certain neoplasms of mesenchymal and glial origin. This notion has been based on observations that these tumors express PDGF genes and produce PDGF-like growth factors. Corresponding data on PDGF receptor expression in sarcoma cell lines is essentially lacking. The cloning of cDNA for 2 distinct PDGF receptors with different abilities to recognize the members of the PDGF family and availability of recombinant PDGF for binding studies have recently made it possible to study the expression of both receptor types in tumor cell lines. We present here a study on 8 human sarcoma cell lines, and show a large variability and independency in the expression of the 2 PDGF receptor types as well as of the genes encoding the corresponding ligands.

Humans↗
PubMed1990

Expression of PDGF beta-receptors in human meningioma cells.

Meningioma is a generally benign tumor derived from arachnoid tissue. We have investigated the presence of functionally active PDGF-receptors on human meningioma cells in culture. Tumor samples were obtained from 3 surgically removed benign meningiomas and normal arachnoid tissue from an autopsy case. Binding studies were performed by using 125I-labelled recombinant PDGF-AA and PDGF-BB. Only 125I-PDGF-BB showed specific binding to all tumor-cell cultures after incubation of cells for 2 hr at 4 degrees C. Effects of PDGF-AA and PDGF-BB on DNA synthesis were measured as 3H-thymidine incorporation during 48 hr of labelling cells maintained in Eagle's minimum essential medium 0.5% fetal calf serum. PDGF-BB but not PDGF-AA stimulated DNA synthesis in all 3 tumor-cell cultures. Total cellular RNA was analyzed by Northern blotting and hybridization with a 32P-labelled human PDGF beta-receptor probe, and PDGF beta-receptor mRNA was found in both tumor and arachnoid cell cultures. Furthermore, PDGF beta-receptor mRNA was shown to be present in 2 meningioma biopsies and immunohistochemical staining revealed that PDGF beta-receptors are present in meningioma and arachnoid tissues in vivo. It appears that a possible way of maintaining human meningioma cell growth in vivo is through activation of PDGF beta-receptors.

Adult↗
PubMed1990

Structural and functional aspects of platelet-derived growth factor and its receptors.

Platelet-derived growth factor (PDGF) is a dimeric molecule that exists as homodimers or heterodimers of related polypeptide chains (A and B). Two types of PDGF receptor have been identified. The PDGF alpha-receptor binds all three isoforms with high affinity whereas the beta-receptor binds only PDGF-BB with high affinity, PDGF-AB with low affinity and does not appear to bind PDGF-AA. The alpha- and beta-receptors are structurally related, each having an intracellular protein tyrosine kinase domain. Ligand-induced functional activation of the receptors appears to involve receptor dimerization. Binding of PDGF to its receptor is followed by internalization and degradation of the ligand-receptor complex. Experiments with mutant receptors have shown that ligand-induced internalization is not absolutely dependent on the kinase activity of the beta-receptor. The v-sis oncogene of simian sarcoma virus (SSV) is a retroviral version of the PDGF B chain gene and SSV-transformation is mediated by an autocrine PDGF-like growth factor. Formal evidence that the expression of the PDGF beta-receptor is sufficient to confer susceptibility to SSV-transformation has been obtained using porcine endothelial cells expressing a recombinant human beta-receptor. PDGF is a chemotactic agent for several cell types. Recent experiments have shown that the PDGF beta-receptor mediates a chemotactic response and that this effect requires an intact protein tyrosine kinase activity.

Animals↗
PubMed1990

Effects of 131I-EGF on cultured human glioma cells.

Malignant glioma cells often have more epidermal growth factor (EGF) receptors than normal cells and targeting of toxic substances to the receptor might therefore be an attractive therapeutical approach. Radiation effects were analysed on human glioma cells growing as monolayers after exposure to 131I-EGF. Unspecific effects were analysed with 131I-BSA or after presaturation with nonradioactive EGF. The radiation effects were compared to the effects obtained by external 60Co gamma irradiation. Administration of the highest radioactive concentrations, 0.2-0.5 MBq/ml in the culture medium, corresponded, after 20 min incubation, to a binding of about 1.0-2.5 dpm/cell. Such an exposure to 131I decays gave effects on cell survival corresponding to about 2.5 Gy of external gamma irradiation. Somewhat less than half of this effect came from the specific bound radioactivity and the rest from nonbound radioactivity. When administrating lower concentrations of radioactivity both the binding and the radiation effects were smaller. The observations showed that it is possible to inactivate cell-proliferation of glioma cells with specific bound 131I-EGF. The possibilities to fractionate the treatments and of binding also other toxic agents than 131I to the EGF receptor are discussed.

Cell Line↗
PubMed1990

Growth inhibition of mitogen-stimulated fibroblasts induced by double-stranded RNA depends on cell density.

Polyinosinic-polycytidylic acid [poly(I:C)], a synthetic double-stranded RNA, is an inhibitor of mitogen-induced proliferation of normal fibroblasts. We show that this inhibition depends strongly on cell density. While cultures with densities at or above confluence are completely inhibited by poly(I:C) in their proliferative response to epidermal growth factor (EGF), the proliferation of sparse (subconfluent) cultures is only delayed. Conditioned medium from dense fibroblasts exposed to poly(I:C) inhibits EGF stimulation of sparse cells, indicating that the inhibition is, at least in part, mediated by a factor released from the cells. Preincubation of quiescent cultures with poly(I:C) renders the cells refractory to the inhibitory effects of poly(I:C). This desensitization correlates with a decreased production of the inhibitor. Since the inhibition of mitogenic stimulation by poly(I:C) is completely overcome by antisera recognizing interferon-beta (IFN-beta) and interleukin-6 (IL-6), we tested the effect of IL-6 and IFN-beta on EGF mitogenicity. None of the available IL-6 preparations had any effect on cell cycle entry. IFN-beta caused a dose-dependent delay of cell division but did not affect the density-dependent proportion of cells entering the cell cycle in response to EGF. Thus, IFN-beta cannot be the sole mediator of the poly(I:C)-induced inhibition. In the presence of dexamethasone, poly(I:C) did not inhibit EGF mitogenis. Indeed, the combined presence of poly(I:C) and dexamethasone did more than just restore the density-dependent control levels of EGF stimulation; most cells entered the cell cycle even at extremely high cell densities. Thus, poly(I:C) in combination with dexamethasone could deactivate the cell density-dependent negative control of proliferation.

Biological Factors↗
PubMed1990

B-type receptor for platelet-derived growth factor mediates a chemotactic response by means of ligand-induced activation of the receptor protein-tyrosine kinase.

Porcine aorta endothelial cells are devoid of receptors for platelet-derived growth factor (PDGF). We have transfected such cells with cDNA for the PDGF B-type receptor, both the wild-type receptor and a mutant form of the receptor (K634A), in which the putative nucleotide-binding lysine of the protein-tyrosine domain has been changed to alanine. Immunoprecipitation studies of metabolically labeled cells showed that both types of receptors were synthesized and processed to the mature form of Mr 190,000. In cells expressing the wild-type receptor, PDGF-BB, the natural ligand for the B-type receptor, induced membrane ruffling and reorganization of actin. Such a response has previously been seen in cells expressing the natural PDGF B-type receptor in response to PDGF-BB. No such effect was induced in nontransfected cells or in cells expressing the K634A mutant receptor. PDGF was also shown to be chemotactic for cells expressing the wild-type receptor, whereas no chemotactic response was elicited in control cells or in cells expressing the K634A mutant receptor. Our study thus provides formal evidence that the PDGF B-type receptor mediates a motility response including actin reorganization and chemotaxis. Furthermore, the results establish a role for the receptor-associated protein-tyrosine kinase in the transduction of the chemotactic signal.

Animals↗
PubMed1990

Differential effects of the various isoforms of platelet-derived growth factor on chemotaxis of fibroblasts, monocytes, and granulocytes.

The chemotactic activities of three different isoforms of platelet-derived growth factor (PDGF) on fibroblasts, monocytes, and granulocytes of human origin were investigated. PDGF-AB and PDGF-BB induced strong, dose-dependent responses in both fibroblasts and monocytes, whereas PDGF-AA did not stimulate chemotaxis of these cell types. Instead, PDGF-AA inhibited the chemotactic activity of PDGF-AB and PDGF-BB on fibroblasts and monocytes. However, PDGF-AA was not able to block monocyte chemotaxis induced by FMLP. In contrast, in granulocytes, dose-dependent chemotactic responses were obtained with all three isoforms of PDGF. All isoforms gave maximal responses at concentrations between 5 and 20 ng/ml. At higher concentrations the migration was reduced. Reduction and alkylation of the PDGF molecule, which leads to loss of the mitogenic activity, also caused a loss of the chemotactic activities for all three cell types. The data suggest that the various isoforms of PDGF stimulate and inhibit chemotaxis in an isoform- and cell type-specific manner.

Alkylation↗
PubMed1990

Localization of platelet-derived growth factor (PDGF) in CHO cells transfected with PDGF A- or B-chain cDNA: retention of PDGF-BB in the endoplasmic reticulum and Golgi complex.

Platelet-derived growth factor (PDGF) is a powerful mitogen for connective tissue cells. It is made up of two polypeptide chains (A and B) and exists in three dimeric forms (AA, AB, and BB). Transfection experiments have indicated that PDGF-AA and -AB are secreted as 30 x 10(3) Mr products, whereas PDGF-BB is processed into a 24 x 10(3) Mr product and remains associated with the cells. Here, CHO cells were transfected with PDGF B- or A-chain cDNA and the intracellular distributions of the respective gene products were compared by indirect immunofluorescence and immunoelectron microscopy, using primary antibodies specific for PDGF B- and A-chain homodimers. PDGF-BB was most conspicuous in stacked Golgi cisternae. It was also found in the endoplasmic reticulum and in lysosomes. Upon treatment of the cells with the microtubule-disruptive drug nocodazole, the Golgi complex was broken up and its stacks of cisternae were dispersed throughout the cytoplasm together with clusters of lysosomes. After this structural disorganization, the concentration of PDGF-BB to the Golgi stacks was even more prominent than before. Weak reactivity for PDGF-AA was detected in the endoplasmic reticulum and groups of vacuoles, both in control and nocodazole-treated cells, whereas Golgi stacks and lysosomes only seldom were positive. The observations suggest that PDGF-BB is processed and retained within the endoplasmic reticulum and Golgi complex. Eventually, it may also be transferred to lysosomes for degradation. In contrast, PDGF-AA is likely to follow a pathway for bulk flow, including rapid passage through the endoplasmic reticulum and Golgi complex, package in secretory vacuoles, and extracellular release by exocytosis.

Animals↗
PubMed1989

Identification and structural analysis of the A type receptor for platelet-derived growth factor. Similarities with the B type receptor.

Binding analyses using 125I-labeled platelet-derived growth factor (PDGF)-AA and PDGF-BB were used to identify a clonal human glioma cell line (U-343 MGa 31L) which expresses the A type but not the B type receptor for PDGF. The glioma cells were devoid of a B type receptor transcript, and immunoprecipitation with an antiserum raised against a B type receptor peptide rendered no signal. Similar analyses using human foreskin fibroblasts, which express both A and B type PDGF receptors, revealed a B type PDGF receptor-specific 5.5-kilobase pair mRNA in a Northern blot experiment, and 160,000 and 180,000 molecular weight components upon immunoprecipitation. A second antiserum, raised against purified porcine PDGF receptor preparations, was reactive with Mr 140,000 and 170,000 components in the U-343 MGa 31L cells, as well as in human fibroblasts. In addition, this antiserum precipitated the Mr 160,000 and 180,000 components from the fibroblasts. Exposure of cells to PDGF-AA, as well as to PDGF-BB, induced an increased rate of degradation of the Mr 170,000 component in the clonal glioma cells and in fibroblasts. The Mr 180,000 component in fibroblasts was degraded only when cells were exposed to PDGF-BB. This allowed the identification of the Mr 170,000 component as the cell surface expressed form of the A type receptor for PDGF. A structural relatedness between the A and B type PDGF receptors was furthermore indicated by similarities in peptide patterns, after limited proteolytic digestion.

Blotting, Northern↗
PubMed1989

PDGF A chain homodimers drive proliferation of bipotential (O-2A) glial progenitor cells in the developing rat optic nerve.

The bipotential glial progenitor cells (O-2A progenitors), which during development of the rat optic nerve give rise to oligodendrocytes and type 2 astrocytes, are stimulated to divide in culture by platelet-derived growth factor (PDGF), and there is evidence that PDGF is important for development of the O-2A cell lineage in vivo. We have visualized PDGF mRNA in the rat optic nerve by in situ hybridization, and its spatial distribution is compatible with the idea that type 1 astrocytes are the major source of PDGF in the nerve. We can detect mRNA encoding the A chain, but not the B chain of PDGF in the brain and optic nerve, suggesting that the major form of PDGF in the central nervous system is a homodimer of A chains (PDGF-AA). PDGF-AA is a more potent mitogen for O-2A progenitor cells than is PDGF-BB, while the reverse is true for human or rat fibroblasts. Fibroblasts display two types of PDGF receptors, type A receptors which bind to all three dimeric isoforms of PDGF, and type B receptors which bind PDGF-BB and PDGF-AB, but have low affinity for PDGF-AA. Our results suggest that O-2A progenitor cells possess predominantly type A receptors, and proliferate during development in response to PDGF-AA secreted by type 1 astrocytes.

Animals↗
PubMed1989

Isoform-specific induction of actin reorganization by platelet-derived growth factor suggests that the functionally active receptor is a dimer.

Human platelet-derived growth factor (PDGF) occurs as three isoforms which are made up of disulfide-bonded A and B chains. The isoforms bind with different affinities to two different but structurally related cell surface receptors. The A type receptor binds all three isoforms (PDGF-AA, PDGF-AB, PDGF-BB) with high affinity, whereas the B type receptor binds PDGF-BB with high affinity, PDGF-AB with lower affinity but does not appear to bind PDGF-AA. We have utilized the differential effects of the three isoforms on actin reorganization and membrane ruffling in human foreskin fibroblasts to probe the idea that ligand-induced receptor dimerization is associated with receptor activation. Actin reorganization was found to be induced only by PDGF-AB and PDGF-BB and is therefore likely to be mediated by the B type receptor. Simultaneous addition of PDGF-AA, or downregulation of the A type receptor blocked the effect of PDGF-AB but not that of PDGF-BB. This is compatible with a model by which PDGF-AB binds to and dimerizes one A and one B type receptor; PDGF-AB therefore requires A type receptors in order to be functionally active at physiological concentrations. In cells with down-regulated A type receptors, high concentrations of PDGF-AB inhibited the effect of PDGF-BB on actin reorganization. We believe that this is due to a monovalent binding of PDGF-AB to the B type receptors which prevents PDGF-BB from dimerizing the receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗