ABSTRACT
Nonhematopoietic malignant cells may express receptors for hematopoietic growth factors and respond to these peptides. The aim of the present study was to investigate whether small cell lung cancer (SCLC) cells may be stimulated to proliferate by hematopoietic growth factors such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3), which are currently used in clinical trials in combination with cytotoxic chemotherapy. We studied two SCLC cell lines, H69 and N417. The effects of GM-CSF and IL-3 were evaluated by studying clonal growth, 3H-thymidine incorporation, BUDR/DNA bivariate flow cytometry, c-myc and N-myc oncogene expression, and myeloid surface markers. Our experiments show that both GM-CSF and IL-3 can increase 3H-thymidine incorporation and cloning efficiency and reduce DNA synthesis time of H69 and, to a lesser extent, N417 cells, supporting the hypothesis that hematopoietic growth factors can stimulate the growth of some malignant nonhematopoietic cells in vitro. Further in vitro and in vivo studies are needed to determine whether clinical trials applying these factors for bone marrow recovery after chemotherapy of solid tumors may be hazardous by potentially stimulating growth of remaining tumor tissue.
Subject(s)
Carcinoma, Small Cell/pathology , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Interleukin-3/pharmacology , Cell Division/drug effects , DNA, Neoplasm/biosynthesis , Gene Expression Regulation, Neoplastic/drug effects , Genes, myc , Humans , In Vitro Techniques , Tumor Cells, CulturedABSTRACT
The detection of human chromosomes in somatic cell hybrids is usually made by chromosomal analysis, Southern blot analysis with human probes, and starch-gel electrophoresis of isoenzymes. We describe here a new, quick, and very efficient method to detect human chromosomes in somatic cell hybrids between human and rodent (rat and mouse) cells. The method is based on the polymerase chain reaction to promote amplification of human DNA, using primers derived from localized genes or DNA fragments from each human chromosome.
Subject(s)
Chromosomes, Human , Hybrid Cells , Animals , DNA, Single-Stranded , Humans , Molecular Sequence Data , Polymerase Chain Reaction , RatsABSTRACT
We established and characterized two cell lines derived from glioblastoma multiforme. Both cell lines exhibited tumor cell morphology and growth kinetics and showed variable expression of glial fibrillary acidic protein (GFAP), S-100, fibronectin and vimentin. Cytofluorimetrical analysis of tumor samples showed a diploid DNA distribution, whereas permanent culture cells evolved to the hyperdiploid DNA content. Karyotype studies revealed cytogenetical abnormalities described in glial tumors including gain of chromosome 7, loss of chromosome 10 and presence of double minutes (DMs). Enhanced expression of Ha-ras and c-myc genes resulted from high p-21 and p-62 levels. The contemporary presence of TGF-alpha and EGF-Rc transcripts suggested an autocrine mechanism in the cell lines growth.
Subject(s)
Chromosome Aberrations , Chromosome Disorders , Glioblastoma/pathology , Cell Division , Cell Line , Chromosome Banding , Chromosome Deletion , Chromosomes, Human, Pair 7 , Culture Techniques/methods , DNA, Neoplasm/analysis , Diploidy , ErbB Receptors/genetics , Genes, myc , Genes, ras , Glial Fibrillary Acidic Protein/analysis , Glioblastoma/genetics , Humans , Karyotyping , Kinetics , Ploidies , Transcription, Genetic , Transforming Growth Factor alpha/geneticsABSTRACT
Various clinical studies have demonstrated that high-dose medroxyprogesterone acetate (HD-MPA) can reduce hematologic toxicity in patients receiving chemotherapy for advanced solid tumors. The underlying mechanism(s) of this action is still unknown. A direct effect of MPA on hemopoietic cells has been postulated, but in vitro studies have given contradictory results. To clarify the biologic activity of MPA on hemopoiesis we have evaluated in vitro growth of pluripotent and committed progenitor cells from bone marrow cells which were preincubated in vitro with various doses of MPA and subsequently treated with or without the S-phase-specific drug arabinoside-cytosine (Ara-C). Four healthy subjects and 8 patients with advanced stage solid tumors with no bone marrow involvement were studied. In our experimental model we did not observe any effect of MPA on Ara-C killing of progenitor cells from either bone marrow mononuclear cells or bone marrow mononuclear cells depleted of T-lymphocytes and adherent cells. These results suggest that MPA does not act directly (or indirectly through the production of cytokines by T-lymphocytes and/or monocytes and macrophages) on bone marrow progenitors. In addition, the supposed mechanism of rendering stem cells less susceptible to the insult of cytotoxic drugs by lowering the number of progenitors in the S-phase has been ruled out by cell kinetic studies.
Subject(s)
Bone Marrow/drug effects , Cytarabine/antagonists & inhibitors , Hematopoietic Stem Cells/drug effects , Medroxyprogesterone/analogs & derivatives , Humans , Medroxyprogesterone/pharmacology , Medroxyprogesterone AcetateABSTRACT
Small clusters of extra large muscle fibres were identified in hindlimb muscles of neonatal mice (strain C57BL/10ScSn). At two days of age they had a significantly greater cross-sectional area than their normal counterparts (P less than 0.01). Fibre typing methods (NADH-tetrazolium reductase, ATPase and phosphorylase) classified them as 2A fast oxidative glycolytic (FOG fibres). The activity of NADH-tetrazolium reductase and the lysosomal enzymes beta-glucuronidase, acid phosphatase and dipeptidyl peptidase II were all elevated in the large fibres. Microsomal aminopeptidase (mAPP), a membrane-bound enzyme, also showed increased activity. The fibres are probably the mouse equivalent of the Wohlfart B fibres of the human fetus, with which comparison is made.
Subject(s)
Animals, Newborn/metabolism , Muscles/enzymology , Aging/metabolism , Animals , Animals, Newborn/anatomy & histology , Endopeptidases/metabolism , Female , Hindlimb/enzymology , Hydrolases/metabolism , Immunoenzyme Techniques , Male , Mice , Mice, Inbred C57BL/anatomy & histology , Mice, Inbred C57BL/metabolism , Muscles/anatomy & histologyABSTRACT
In an attempt to clarify the role of beta interferon in vitro cell systems, the metabolic expression of dihydrofolate reductase (DHFR), succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH), by histochemical methods was studied. DHFR was also quantified by flow cytometry. Glioblastoma cell line with or without beta interferon was used. Three days after the treatment the DHFR reaction was much less intense than in the control. Quantitative data confirmed these results. Immediately afterwards, most cells exhibited much more intense reaction. These facts underlined that, in this biological model in vitro, beta interferon could reduce the synthesis of these enzymes only for a short period.
Subject(s)
Brain Neoplasms/pathology , Glioma/pathology , Interferon Type I/pharmacology , Neoplasm Proteins/analysis , Oxidoreductases/analysis , Brain Neoplasms/enzymology , Cell Line , Depression, Chemical , Enzyme Induction/drug effects , Glioma/enzymology , Humans , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/enzymologyABSTRACT
A case is reported of bilateral osteoarthritis of the hips treated in successive years by total prosthetic replacement of the Müller type. Six years after the second replacement, having been completely free from symptoms, she developed pain in the left hip due to rupture of the acetabular cup - a very rare complication.
