Autoradiographic localization of tritiated suramin in polarized human colon adenocarcinoma cells.
ABSTRACT In this report we demonstrated that [3H]suramin enters polarized human colon adenocarcinoma cells when added to the apical side of the monolayer. Using light microscopic quantitative autoradiography, we showed that suramin was accumulated in the apical cytoplasm and in the nucleus. In contrast, a weak labeling was noted in other compartments such as the basolateral cytoplasm and the intercellular space. The accumulation of suramin in the apical region of the cells is consistent with previous data showing that suramin elicited a lysosomal storage disorder in HT29-D4 cells by a mechanism of polarized endocytosis.
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ABSTRACT: Suramin, a polyanionic drug used in the treatment of Rhodesian and Gambian trypanosomiasis and more recently in the acquired immune deficiency syndrome, is a potent inhibitor of the constitutive mammalian DNA polymerases alpha, beta, and gamma and the lymphoid-specific polymerase terminal deoxynucleotidyl transferase. To define the effect of this inhibition on cell proliferation, we studied the effect of suramin on several cell lines in culture and in mice in vivo. Suramin, at 200 micrograms/ml (which is regularly achieved in the plasma of patients), had no effect on the proliferation of 4 of 5 nonlymphoid cell lines. In contrast, exposure of 10 lymphoid cell lines to 200 micrograms/ml suramin for 4 days caused significant growth inhibition in 8 of these 10 lines. Suramin given i.p. to BALB/cBYJ mice at clinically relevant doses (15-60 mg/kg) caused profound and prolonged thymic atrophy within 5-7 days of drug administration (greater than a 90% weight loss in mice treated with 60 mg/kg). Thymic sections revealed severe cortical loss, prominence of dendritic cells, and vacuolated macrophages. Liver, peripheral blood, spleen, kidney, and total body weights were not affected. The apparent selective lymphocytotoxicity of suramin may represent an important property of this drug. We speculate that this may account for the persistent immune suppression reported in suramin-treated acquired immune deficiency syndrome patients.Cancer Research 10/1987; 47(17):4694-8. · 8.65 Impact Factor
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ABSTRACT: Monolayer cultures of a rat hepatocyte cell line shown previously to accumulate a nuclear pool of free heparan sulfate chains that are enriched in sulfated glucuronic acid (GlcA) residues (Fedarko, N.S., and Conrad, H.E., (1986) J. Cell Biol. 587-599) were incubated with 35SO4(2-), and the rate of appearance of heparan [35S]sulfate in the nuclei was measured. Heparan [35S]sulfate began to accumulate in the nuclei 2 h after the administration of 35SO4(2-) to the cells and reached a steady state level after 20 h. Heparan [35S]sulfate was lost from the nuclei of prelabeled cells with a t1/2 of 8 h. Chloroquine did not inhibit the transport of heparan sulfate into the nucleus, but increased the t1/2 for the exit of heparan sulfate from the nucleus to 20 h and led to a doubling of the steady state level of nuclear heparan sulfate. Heparan [35S]sulfate which was obtained from the medium or from the cell matrix of a labeled culture and which contained only low levels of GlcA-2-SO4 residues was incubated with cultures of unlabeled cells, and the uptake of the exogenous heparan [35S]sulfate was studied. At 37 degrees C the cells took up proteoheparan [35S]sulfate and transported about 10% of the internalized heparan [35S]sulfate into the nucleus, where it appeared as free chains. The heparan [35S]sulfate isolated from the nucleus was enriched in GlcA-2-SO4 residues, whereas the heparan [35S]sulfate remaining in the rest of the intracellular pool showed a corresponding depletion in GlcA-2-SO4 residues. At 16 degrees C, where endocytosed materials do not enter the lysosomes, the cells also transported exogenous proteoheparan [35S]sulfate to the nucleus with similar processing. Thus, the metabolism of exogenous heparan sulfate by hepatocytes follows the same pathway observed in continuously labeled cells and does not involve lysosomal processing of the internalized heparan sulfate.Journal of Biological Chemistry 11/1986; 261(29):13575-80. · 4.65 Impact Factor
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ABSTRACT: When NIH 3T3 cells were transfected with the cDNA for basic fibroblast growth factor (bFGF), most cells displayed a transformed phenotype. Acquisition of a transformed phenotype was correlated with the expression of high levels of bFGF (Quarto et al., 1989). Cells that had been transformed as a result of transfection with bFGF cDNA had a decreased capacity to bind 125I-bFGF to high affinity receptors. NIH 3T3 cells transfected with bFGF cDNA that expressed lower levels of bFGF were not transformed and had a normal number of bFGF receptors. NIH 3T3 cells transfected with the hst/Kfgf oncogene, which encodes a secreted molecule with 45% homology to bFGF, also displayed a transformed phenotype and decreased numbers of bFGF receptors. However, NIH 3T3 cells transfected with the H-ras oncogene were transformed but had a normal number of bFGF receptors. Thus, transformation by bFGF-like molecules resulted in downregulation of bFGF receptors. Receptor number was not affected by cell density for both parental NIH 3T3 cells and transformed cells. In the cells transfected with bFGF cDNA that were not transformed, the receptors could be downregulated in response to exogenous bFGF. Conditioned medium from transformed transfected cells contained sufficient quantities of bFGF to downregulate bFGF receptors on parental NIH 3T3 cells. Thus, the downregulation of bFGF receptors seemed related to the presence of bFGF in an extracytoplasmic compartment. Treatment of the transformed transfected NIH 3T3 cells with suramin, which blocks the interaction of bFGF with its receptor, reversed the morphological transformation and restored receptors almost to normal numbers. These results demonstrate that in these cells bFGF transforms cells by interacting with its receptor and that bFGF and hst/K-fgf may use the same receptor.The Journal of Cell Biology 12/1989; 109(5):2519-27. · 10.82 Impact Factor