sepharose and Carcinoma

Topics: Aged; Ascitic Fluid; Carcinoma; Female; Histocytological Preparation Techniques; Humans; Immunohistochemistry; Ovarian Neoplasms; Sepharose

Imbalances in the epithelial-stromal interactions are important in the pathogenesis of prostate cancer. However, we know little about androgenic regulation in the stroma of prostate cancer. We examined the cancer-stromal interaction paying attention to androgen responsiveness of stromal side. In co-culture, PC3 and LNCaP cells did not affect dihydrotestosterone (DHT)-dependent growth of prostate stromal cells (PrSCs), but DU145 cells significantly reduced it. Conditioned medium from DU145 cells (DU145-CM) also inhibited DHT-dependent PrSCs growth, androgen receptor (AR) expression, and prostate specific antigen transcription. Although the inhibitory effect of DU145-CM was not affected by neutralizing antibody against EGF, FGF-2, or TNF-alpha, pretreatment with testosterone-Sepharose partially reduced the inhibitory ability of DU145-CM. These results suggest that DU145 cells produce inhibitory factors for androgen responsiveness, including steroid-binding protein(s), and these may participate in crosstalk between DU145 cells and PrSCs as modulators of androgen.

Topics: Carcinoma; Coculture Techniques; Culture Media, Conditioned; Dihydrotestosterone; Epidermal Growth Factor; Fibroblast Growth Factor 2; Humans; Male; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Receptors, Androgen; Sepharose; Stromal Cells; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

The efficacy of agarose in preventing VX2 carcinoma cell leakage was evaluated and the results were compared with two traditional methods. Forty-five rabbits were divided into 3 groups: Group 1, VX2 tumor cells were injected directly into the liver and no special procedure after removal of the needle; Group 2, the puncture site was gently compressed, using an alcoholic cotton gauze, for three minutes; Group 3, 0.2 ml of heated liquid agarose was injected to seal the aperture after injection of VX2 cells. The leakage rates were 80%, 53.3% and 6.6% for group 1, group 2 and group 3, respectively. We consider agarose is a useful material in preventing the leakage in the establishment of VX2 liver tumor models.

Topics: Animals; Carcinoma; Disease Models, Animal; Hot Temperature; Injections; Liver Neoplasms, Experimental; Neoplasm Seeding; Neoplasm Transplantation; Rabbits; Sepharose

We studied the influence of 10 synthetic glycoamine analogs on colony formation in 0.3 and 0.9% agarose by metastatic human breast carcinoma (MDA-MB-435) and melanoma (TXM-13) cells. Nine synthetic analogs significantly inhibited the colony formation in 0.9% agarose of MDA-MB-435 human breast carcinoma cells; five compounds caused a 73-83% reduction of colony formation. Seven synthetic glycoamines caused a significant inhibition of colony formation in 0.9% agarose by TXM-13 melanoma cells with the inhibitory effect ranging from 71 to 87%. The 50% inhibition (I50) doses and relative activity rank of the compounds were similar for both breast carcinoma and melanoma cell lines. The murine B16 melanoma cell aggregation assay was employed to elucidate the potential mechanism(s) of the inhibitory activity of synthetic glycoamines. The relative activity ranks of the compounds based on the independently determined I50 doses for both cell aggregation and clonogenic growth assays were very similar for the four most active synthetic analogs and clearly indicated the importance of hydrophobic amino acid in mediating the bioactivity of synthetic glycoamines. In both experimental systems (clonogenic growth in agarose and cell aggregation assay) the leading compound was N-(1-deoxy-D-fructos-1-yl)-D-leucine (Fru-D-Leu) and the least active analog was N-(l-deoxy-D-fructos-1-yl)-glycine (Fru-Gly). These results show that synthetic glycoamines may act by competing for specific carbohydrate-lectin interactions, particularly those involving beta-galactoside-specific lectins expressed on metastatic cells.

Topics: Amino Sugars; Animals; Binding, Competitive; Breast Neoplasms; Carbohydrate Sequence; Carcinoma; Colony-Forming Units Assay; Female; Galactosides; Glucosamine; Humans; Lectins; Magnetic Resonance Spectroscopy; Mass Spectrometry; Melanoma; Mice; Molecular Sequence Data; Molecular Structure; Neoplasm Metastasis; Sepharose; Structure-Activity Relationship; Tumor Cells, Cultured; Tumor Stem Cell Assay

We determined the optimal conditions for the separation of N-glycosylation variants of prostate-specific antigen using concanavalin A. Concanavalin A is a lectin that binds to the terminal sugar residues of glycoproteins. We demonstrated that differences in the percentage of prostate-specific antigen bound to concanavalin A-Sepharose in patients with benign prostatic hyperplasia compared with patients with prostatic carcinoma, as described in the literature, arise when insufficient concanavalin A binding sites are added for complete binding of the glycosylation variants of prostate-specific antigen. We observed similar percentages of prostate-specific antigen bound to concanavalin A-Sepharose for benign prostatic hyperplasia (86.3% +/- 7.5, mean +/- SD) and carcinoma patients (81.8% +/- 12.0, mean +/- SD), when sufficient concanavalin A-Sepharose was added to allow optimal binding, and when samples with high prostate-specific antigen concentrations were not pre-diluted before incubation with concanavalin A-Sepharose. We conclude that differentiation of patients with benign prostatic hyperplasia or carcinoma of the prostate on the basis of differences in percentages of prostate-specific antigen bound to concanavalin A-Sepharose, i.e. separation of N-glycosylation variants, is not possible.

Topics: Binding Sites; Carcinoma; Chromatography, Affinity; Concanavalin A; Glycosylation; Humans; Male; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms; Sepharose

Differences in tumorigenicity between the Epstein-Barr virus (EBV) genome-negative and -positive epithelial A2L/AH hybrid cell lines have been described recently. Further differences in growth characteristics of the two epithelial hybrid cells have since been examined. Differences were found in doubling time and cloning efficiency in agarose of the two cell lines. In addition, the EBV genome-negative epithelial hybrid cell line (cl-654), unlike the EBV genome-positive epithelial hybrid cell line (cl-1), failed to grow in the unmanipulated nude mice, but was successfully transplanted in 60Co-irradiated nude mice. The histopathologic findings of tumors induced by the inoculation of cl-654 cells in nude mice showed more differentiated carcinoma than the findings of tumors induced by cl-1 cells. The data suggest that EBV is associated with an alteration of cell differentiation in epithelial cells as well as tumorigenicity in nude mice.

Topics: Animals; Carcinoma; Cell Division; Cell Line; Cell Transformation, Neoplastic; Clone Cells; Culture Media; Epithelial Cells; Female; Genes, Viral; Herpesvirus 4, Human; Hybrid Cells; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Sepharose

Affinity chromatography on a concanavalin A (con A)-Sepharose column is a potentially useful for the isolation of whole thyroglobulin (Tg) at least from normal thyroid tissue. In addition to being a simple procedure for the isolation of Tg, large amounts of Tg can be applied to the column and recovered in good yield with a buffer containing MeG. In gradient elution with buffer containing increasing amounts of MeG, a single but broad peak was obtained, without separation into subfractions. However, a hemagglutination-inhibition test showed that the Tg preparation eluted early from the column had less affinity for con A than the Tg preparation eluted later, suggesting a heterogeneous distribution of carbohydrate moieties among Tg preparations. When human Tg from thyroid tumor was applied to the column, tumor Tg partly passed through the column without being adsorbed. This unadsorbed Tg showed a very low affinity for lectins, con A and wheat germ agglutinin (WGA), as determined by a double diffusion reaction in agar gel. In contrast to this fraction, the Tg adsorbed on the con A-gel column showed a very strong affinity for WGA, differing from Tg of normal thyroid tissue. Therefore, tumor Tg preparation appears to have an abnormally modified carbohydrate structure, at least in part. The higher affinity for WGA (with a specificity for N-acetylglucosamine) seen in adsorbed Tg could be due to a larger amount of GlcNAc residues which bind irregularly in the carbohydrate moiety of tumor Tg.

Topics: Adenoma; Animals; Carcinoma; Chromatography, Affinity; Humans; Immunodiffusion; Male; Orchiectomy; Reference Values; Sepharose; Swine; Thyroglobulin; Thyroid Gland; Thyroid Neoplasms