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21.
Metabolically 35S-labeled proteoglycans were isolated from cell-associated matrices and media of confluent cultures of human normal transitional epithelial cells and HCV-29T transitional carcinoma cells. On Sepharose CL-4B columns, the cell-associated proteoglycans synthesized from both cell types separated into three identical size classes, termed CI, CII, and CIII. Normal epithelial cell C-fractions eluted in a 22:34:45 proportion and contained 64%, 64%, and 72% heparan sulfate, whereas corresponding HCV-29T fractions eluted in a 29:11:60 proportion, and contained 91%, 77%, and 70% heparan sulfate, respectively. Medium proteoglycans from normal cells separated into two size classes in a proportion of 6:94 and were composed of 35% and 50% heparan sulfate. HCV-29T medium contained only one size class of proteoglycans consisting of 23% heparan sulfate. The remaining percentages were accounted for by chondroitin/dermatan sulfate. On isopycnic CsCl gradients, proteoglycan fractions from normal cells had buoyant densities that were higher than the corresponding fractions from HCV-29T cells. DEAE-Sephacel chromatography showed that cell and medium associated heparan sulfate from HCV-29T cells was consistently of lower charge density (undersulfated) than that from normal epithelial cells. In contrast, the chondroitin/dermatan sulfate of HCV-29T was of a charge density similar to that of normal cells. These as well as other structural and compositional differences in the proteoglycan may account, at least in part, for the altered behavioral traits of highly invasive carcinoma cells. 相似文献
22.
Jamal Bayad Nicole Sabolovic Denyse Bagrel Athanase Visvikis Maria Wellman Gerard Siest 《Cell biology and toxicology》1990,6(2):157-170
Eighteen IgGl monoclonal antibodies (blabs) have been produced against gamma-glutamyl transferase (GGT) from rat kidney. They were specific to the light subunit of the enzyme with affinity constants ranging from 0.3 to 7.5 108 M–1, while they did not react with GGT from other sources i.e. human and pig kidney, rat and guinea pig liver, suggesting species and organ specificity. Two of the blabs (N° 11 and 21) lost their immunoreactivities towards rat kidney GGT in the presence of N-acetyl-neuraminic acid, while immunoreactivities of the other blabs were unchanged. Furthermore, Mabs No 11 and 21 did not react with desialylated rat kidney GGT. These findings suggest that N-acetyl-neuraminic acid is involved in the epitopes recognized by these two Mabs.Abbreviations ELISA
enzyme linked immunosorbent assay
- GGT
gamma-glutamyltransferase
- Mab
monoclonal antibody
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
23.
D.W. Bruno 《FEMS microbiology letters》1988,51(2-3):135-139
Abstract The cell surface hydrophobicity of Renibacterium salmoninarum strains was examined using a salt aggregation method. Those strains which were virulent in the test animal were sticky, auto-agglutinating and possessed a hydrophobic cell surface. Those strains with a low virulence were non-sticky, non-agglutinating and failed to aggregate in a high molar salt. Strains could not be distinguished using biochemical tests. There was no change in hydrophobicity following re-isolation of the bacteria from experimentally infected rainbow trout, Salmo gairdneri . 相似文献
24.
Agnes Janoshazi David M. Ojcius Bruce Kone Julian L. Seifter A. K. Solomon 《The Journal of membrane biology》1988,103(2):181-189
Summary A membrane protein that is immunochemically similar to the red cell anion exchange protein, band 3, has been identified on the basolateral face of the outer medullary collecting duct (MCD) cells in rabbit kidney. In freshly prepared separated rabbit MCD cells, M.L. Zeidel, P. Silva and J.L. Seifter (J. Clin. Invest.
77:1682–1688, 1986) found that Cl–/HCO
3
-
exchange was inhibited by the stilbene anion exchange inhibitor, DIDS (4,4-diisothiocyano-2,2-disulfonic stilbene), with aK
1 similar to that for the red cell. We have measured the binding affinities of a fluorescent stilbene inhibitor, DBDS (4,4-dibenzamido-2,2-disulfonic stilbene), to MCD cells in 28.5 mM citrate and have characterized both a high-affinity site (K
1
s
=93±24 mM) and a lower affinity site (K
2
s
=430±260 nM), which are closely similar to values for the red cell of 110±51 nM for the high-affinity site and 980±200 nM for the lower affinity site (A.S. Verkman, J.A. Dix & A.K. Solomon,J. Gen. Physiol.
81:421–449, 1983). When Cl– replaces citrate in the buffer, the two sites collapse into a single one withK
1
s
=1500±400 nM, similar to the singleK
1
s
=1200±200 nM in the red cell (J.A. Dix, A.S. Verkman & A.K. Solomon,J. Membrane Biol.
89:211–223, 1986). The kinetics of DBDS binding to MCD cells at 0.25 M–1 are characterized by a fast process, =0.14±0.03 sec, similar to =0.12±0.03 sec in the red cell. These similarities show that the physical chemical characteristics of stilbene inhibitor binding to MCD cell band 3 closely resemble those for red cell band 3, which suggests that the molecular structure is highly conserved. 相似文献
25.
The metabolism and mutagenic activation of 2-acetylaminofluorene by human and rat hepatocytes and kidney cells were measured. High performance liquid chromatography was used to separate the 2-acetylaminofluorene metabolites, and a cell-mediated Salmonella typhimurium mutagenesis assay was used to detect mutagenic intermediates. Rat and human differences were observed with cells from both organs and levels of metabolism and mutagenesis were higher in human cells. Within a species, liver and kidney cell differences were also evident, with levels of hepatocyte-mediated metabolism and mutagenesis being greater than kidney cells. Human inter-individual variation was apparent with cells from both organs, but the variation observed was significantly greater in hepatocytes than kidney cells. A knowledge of such differences, including an understanding that they may vary with the chemical being studied, should be useful in the extrapolation of rodent carcinogenesis data to humans.Abbreviations AAF
2-acetylaminofluorene
- AF
2-aminofluorene
- DMSO
dimethylsulfoxide
- HPLC
high performance liquid chromatography
- N-OH-AAF
N-hydroxy-2-acetylaminofluorene
- 1-OH-AAF
1-hydroxy-2-acetylaminofluorene
- 3-OH-AAF
3-hydroxy-2-acetylaminofluorene
- 5/9-OH-AAF
a combination of 5 and 9-hydroxy-2-acetylaminofluorene
- 7-OH-AAF
7-hydroxy-2-acetylaminofluorene
- 8-OH-AAF
8-hydroxy-2-acetylaminofluorene 相似文献
26.
27.
小鼠Lewis肺癌组织中层粘连蛋白受体的分离及其性质的研究 总被引:2,自引:2,他引:0
层粘连蛋白(Laminin,LN)是基膜(basement membrane)中的一种主要大分子糖蛋白。一些研究资料表明肿瘤细胞的浸润转移可能与LN有关。肿瘤细胞与LN的作用可能是通过细胞表面LN受体进行的。本文采用亲和层析法从小鼠Lewis肺癌组织中分离LN受体并对其理化性质进行研究。Lewis肺癌LN受体的表观分子量为70,000,还原后SDS电泳图为一条较宽的条带。氨基酸组成中疏水氨基酸占38%,苏氨酸、絲氨酸、门冬氨酸(包括门冬酰胺)占23.5%,通过硝酸纤维素膜片法用HRP-LN测定受体与LN的结合特性,证明具有配基结合专一性,饱和性及高亲和性(Kd=0.95×10~(-9)mol/L)。 相似文献
28.
Paul H. Gumerlock Benjamin F. Edwards Arline D. Deitch Frederick J. Meyers 《In vitro cellular & developmental biology. Plant》1988,24(5):429-434
Summary A human cell line has been established from a renal adenocarcinoma rib metastasis of a 58-y-old male. This cell line has been
maintained in continuous culture for 20 mo. through more than 50 passages. It displays simulataneous expression of the intermediate
filaments cytokeratin and vimentin. Flow cytometric analysis of DNA content reveals a major hyperdiploid population.
This work was supported in part by a grant from Triton Biosciences, Inc. 相似文献
29.
Adriaan P. de Bruïne Winand N. M. Dinjens Margriet M. J. Pijls Edith P. M. v. d. Linden Mat J. M. Rousch Peter T. Moerkerk Antony F. P. M. de Goeij Fred T. Bosnian 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,62(1):311-320
In colonic neoplasms, endocrine differentiation is encountered not only in carcinoid tumors but also in adenocarcinomas, where
endocrine cells may represent a distinct line of differentiation in the tumor. The significance of endocrine differentiation
in colorectal cancer is not well established, partly because of the paucity of tumor cell lines which can serve as a model
for studying endocrine differentiation. In this report we describe the properties of NCI-H716 cells, a cell line derived from
a poorly differentiated adenocarcinoma of the caecum, under various in vitro conditions and as xenografts in athymic mice.
Phenotypical properties were immunohistochemically assessed using a panel of differentiation related antibodies, and also
by Northern blot analysis and by electron microscopy. Receptors for biogenic amines and peptide hormones were analyzed by
ligand binding assay. These studies show that:
相似文献
1. | NCI-H716 cells can be undifferentiated, or show endocrine, mucin-producing or “amphicrine” properties. |
2. | Endocrine differentiation of NCI-H716 cells preferentially occurs in xenografts in athymic mice, which suggests that mesenchymal elements induce endocrine differentiation. |
3. | NCI-H716 cells express large amounts of high affinity receptors for gastrin, serotonin and somatostatin and these substances can regulate growth. Thus, NCI-H716 cells form a suitable model for the study of endocrine differentiation in intestinal epithelium and of auto- or paracrine growth regulation in intestinal neoplasia. |
30.
Daniel R. Vlock Beth Arnold Jeannette Humpierres Donald R. Schwartz Shan R. Baker Charles J. Krause Neil Swanson Thomas E. Carey 《Cancer immunology, immunotherapy : CII》1992,34(5):329-336
Summary In previous studies we evaluated the incidence and specificity of autologous antibody reactivity against squamous cell carcinoma of the head and neck (SCCHN). We were able to demonstrate that autologous antibody reactivity is present in native sera but was usually of too low a titer to allow further analysis. Dissociation of immune complexes by acidification and ultrafiltration of serum augmented autologous antibody reactivity in nine out of nine autologous systems tested. Native antibody and antibody derived from immune complexes produced by the host and reactive with autologous tumor cells may be directed against physiologically relevant antigens. Therefore, correlations of antibody titers with clinical course may provide insight into the nature of the host response to cancer. In the present analysis, serological studies of six patients with SCCHN were performed with serum samples obtained over many months. Results of serial serological assays were correlated to tumor progression and clinical course. Fluctuations in autologous antibody reactivity were noted over time. In four cases, rises in autologous antibody titers preceded the clinical diagnosis of recurrence by several months. Drops in autologous antibody reactivity were noted in two cases following surgery or radiation therapy. In two cases of long-term survivors, no correlation between antibody reactivity and clinical course was noted. Specificity analysis of the six autologous systems demonstrated reactivity against autologous and allogeneic SCCHN as well as melanoma cell lines. These sera did not react with glioma, neuroblastoma, renal cell, breast, bladder and colon carcinoma cell lines nor with fetal calf serum, pooled lymphocytes, red blood cells and platelets. Autologous serial serological studies may provide a means by which to evaluate the host/tumor relationship in patients with SCCHN. 相似文献