concanavalin-a and Prostatic-Neoplasms

Prostate-specific antigen (PSA) is the most frequently used biomarker for the screening of prostate cancer. Understanding the structure of cancer-specific glycans can help us improve PSA assay. In the present study, we analysed the glycans of PSA obtained from culture medium containing cancer tissue-originated spheroids (CTOS) which have similar characteristics as that of the parent tumour to explore the new candidates for cancer-related glycoforms of PSA. The glycan profile of PSA from CTOS was determined by comparing with PSA from normal seminal plasma and cancer cell lines (LNCaP and 22Rv1) using lectin chromatography and mass spectrometry. PSA from CTOS was mostly sialylated and the content of Wisteria floribunda agglutinin reactive glycan (LacdiNAc) was similar to that of PSA derived from seminal plasma and 22Rv1. Conversely, concanavalin A (Con A)-unbound PSA was definitely detected from the three cancer origins but was almost negligible in seminal PSA. Two novel types of PSA were elucidated in the Con A-unbound fraction: one is a high molecular weight PSA with highly branched N-glycans, and the other is a low molecular weight PSA without N-glycans. Furthermore, the existence of Lewis X antigen group on PSA was indicated. These PSAs will be candidates for new cancer-related markers.

Topics: Biomarkers, Tumor; Carbohydrate Sequence; Cell Line, Tumor; Chromatography, Affinity; Concanavalin A; Culture Media, Conditioned; Glycopeptides; Glycosylation; Humans; Lewis X Antigen; Male; Plant Lectins; Polysaccharides; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Protein Processing, Post-Translational; Receptors, N-Acetylglucosamine; Semen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spheroids, Cellular

Lectins are carbohydrate recognition proteins that can be used as probes to reveal the glycosylation state of cells. They frequently have been used for diagnostic and prognostic cancer studies. For fluorescence based analysis, lectins commonly are conjugated to fluorescein isothiocyanate (Con A-FITC); however, this molecule loses its fluorescence quickly. We conjugated Europium cryptate to Con A (Con A-cryp-Eu) for use as a histochemical luminescent probe to recognize glucose/mannose residues in benign prostatic hyperplasia and prostatic carcinoma tissues, and used confocal microscopy instead of commercial Con A-FITC. Tissues were treated with Evans blue to suppress intrinsic tissue fluorescence before incubation with Con A-cryp-Eu or Con A-FITC. Con A-cryp-Eu exhibited hemagglutinating activity. Con A-cryp-Eu showed the same binding pattern as Con A-FITC in prostate stroma and gland cells. Staining was strong in benign prostate hyperplasia and prostate carcinoma tissues. Con A-cryp-Eu probe stained glucose/mannose residues in prostatic carcinoma more intensely than Con A-FITC. Furthermore, staining with Con A-cryp-Eu showed greater fluorescence intensity than Con A-FITC and the emission of Con A-cryp-Eu was more stable than the Con A-FITC for seven days under the same storage conditions. Maintenance of the luminescent properties and the binding pattern of Con A-cryp-Eu favor its use as an auxiliary histochemistry probe for prostatic tissue studies.

Topics: Concanavalin A; Humans; Luminescent Agents; Male; Microscopy, Confocal; Organometallic Compounds; Prostatic Neoplasms; Sensitivity and Specificity

In this study, we report the development of a microbore hollow fiber enzyme reactor (mHFER) coupled to nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS/MS) for the online digestion or selective enrichment of glycopeptides and analysis of proteins. With mHFER, enzymatic digestion of protein could be achieved by continuous flow within a very small volume (~10 μL) of mHF inserted in a PEEK tube. Digested peptides exited through the pores of the hollow fiber membrane wall to external single or multiplexed trap columns for nLC-ESI-MS/MS analysis. Evaluation of online mHFER-nLC-ESI-MS/MS system was made with bovine serum albumin (BSA) by varying the temperature of digestion and the amount of protein injected. We evaluated the ability of the mHFER system to enrich glycopeptides by injecting a mixture of lectin (concanavalin A) and digested peptides from α-1-acid glycoprotein (AGP) into the mHFER, followed by delivery of PNGase F for endoglycosidic digestion. Nonglycosylated peptides unbound to lectins eluted at the first breakthrough run while N-linked glycopeptides eluted after the endoglycosidic digestion. The developed method was applied to urine samples from patients with prostate cancer and controls; 67 N-linked glycopeptides were identified and relative differences in glycopeptide content between patient and control samples were determined.

Topics: Adipokines; Bioreactors; Carrier Proteins; Case-Control Studies; Chromatography, Liquid; Concanavalin A; Glycopeptides; Glycoproteins; Humans; Male; Orosomucoid; Peptide Fragments; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Prostate; Prostatic Neoplasms; Proteomics; Serum Albumin, Bovine; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Lectins, proteins which selectively recognize carbohydrates, have been used in histochemistry for the evaluation of changes in glycosylation in processes of cellular differentiation and/or dedifferentiation. Cratylia mollis seed lectins (Cramoll 1,4 and Cramoll 3), conjugated to horseradish peroxidase, were used as histochemical probes in human prostate tissues: normal (NP), hyperplasia (BPH), and prostate carcinoma (PCa). The staining pattern of Con-A and Cramoll 1,4 in BPH was more intense than in NP. These lectins also showed staining differences between BPH and PCa; the latter showing decreased staining intensity with an increased degree of malignancy. PNA and Cramoll 3 stained epithelial cells similarly in all diagnoses although they did present intense staining of PCa glands lumen. Corpora amylacea were not differentially recognized by any of the lectins. Cramoll 1,4 and Cramoll 3 seed lectins present themselves as candidates for histochemical probes for prostate pathologies when compared to commercial lectins such as Con-A and PNA.

Topics: Adult; Aged; Aged, 80 and over; Concanavalin A; Fabaceae; Glycosylation; Histocytochemistry; Humans; Male; Middle Aged; Peanut Agglutinin; Plant Lectins; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Seeds; Statistics, Nonparametric

We investigated the effects of the recently synthetized NO donating agent GIT-27NO on the growth of human androgen independent and androgen dependent PC3 and LnCap cells xenografted in nude mice. We also tested the effects of GIT-27NO in the preclinical model of cell-mediated immunoinflammatory hepatitis that can be induced in mice by Concanavalin A (ConA) and that has been shown to benefit from the treatment with NO donating agents such as NO-aspirin. In agreement with in vitro data showing dose-dependent reduction of PC3 and LnCap cell viability with GIT-27NO, the i.p. treatment of mice xenografted with either of these cells with GIT-27NO significantly inhibited their growth as compared to the mice-treated with its vehicle. In addition, GIT-27NO given -24 and -1 h prior to e.v. challenge with 20 mg/kg ConA significantly suppressed the increase of transaminases that occurred 8 h after challenge in the control mice that received the vehicle. In addition, relative to these latter groups of mice, the histological signs of inflammatory hepatitis were markedly reduced in ConA-challenged mice that received GIT-27NO. In the hepatitis model, GIT-27NO was equally effective in preventing ConA-induced hepatitis regardless of whether it was administered intra peritoneally or per os. These data confirm that GIT-27NO is a powerful anticancer agent also endowed with pharmacological properties to prevent the development of cell-mediated murine immunoinflammatory hepatitis.

Topics: Acetates; Animals; Antineoplastic Agents; Cell Line, Tumor; Cell Survival; Concanavalin A; Female; Hepatitis, Autoimmune; Humans; In Vitro Techniques; Male; Mice; Mice, Inbred Strains; Mice, Nude; Neoplasm Transplantation; Nitric Oxide Donors; Oxazoles; Prostatic Neoplasms; Transplantation, Heterologous

As a continuation of our work on boronic acid lectin affinity chromatography (BLAC), in this paper we introduce an automated affinity micropartitioning approach using combined boronic acid and concanavalin A (BLAC/Con A) resin-filled micropipette tips to isolate and enrich human serum glycoproteins. The N-linked oligosaccharides of the partitioned glycoproteins were removed by PNGase F enzyme digestion, followed by 8-aminopyrene-1,3,6-trisulfonic acid labeling. Capillary gel electrophoresis with blue LED-induced fluorescence detection was applied in a multiplexed format for comparative glycan profiling. The efficiency of BLAC affinity micropartitioning was compared with that of the individual lectin and pseudolectin affinity enrichment. Finally, we report on our findings in glycosylation differences in human serum samples from healthy and prostate cancer patients by applying BLAC/Con A micropipette tip-based enrichment and comparative multicapillary gel electrophoresis analysis of the released and labeled glycans.

Topics: Boronic Acids; Chromatography, Affinity; Concanavalin A; Electrophoresis, Capillary; Glycoproteins; Glycosylation; Hemofiltration; Humans; Male; Oligosaccharides; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Polysaccharides; Prostatic Neoplasms; Pyrenes; Wheat Germ Agglutinins

Combinations of cellular immune-based therapies with chemotherapy and other antitumour agents may be of significant clinical benefit in the treatment of many forms of cancer. Gamma delta (gammadelta) T cells are of particular interest for use in such combined therapies due to their potent antitumour cytotoxicity and relative ease of generation in vitro. Here, we demonstrate high levels of cytotoxicity against solid tumour-derived cell lines with combination treatment utilizing Vgamma9Vdelta2 T cells, chemotherapeutic agents and the bisphosphonate, zoledronate. Pre-treatment with low concentrations of chemotherapeutic agents or zoledronate sensitized tumour cells to rapid killing by Vgamma9Vdelta2 T cells with levels of cytotoxicity approaching 90%. In addition, zoledronate enhanced the chemotherapy-induced sensitization of tumour cells to Vgamma9Vdelta2 T cell cytotoxicity resulting in almost 100% lysis of tumour targets in some cases. Vgamma9Vdelta2 T cell cytotoxicity was mediated by perforin following TCR-dependent and isoprenoid-mediated recognition of tumour cells. Production of IFN-gamma by Vgamma9Vdelta2 T cells was also induced after exposure to sensitized targets. We conclude that administration of Vgamma9Vdelta2 T cells at suitable intervals after chemotherapy and zoledronate may substantially increase antitumour activities in a range of malignancies.

Topics: Adenocarcinoma; Antineoplastic Agents; Apoptosis; Breast Neoplasms; Burkitt Lymphoma; Carcinoma; Cell Line, Tumor; Cisplatin; Colorectal Neoplasms; Concanavalin A; Cytotoxicity, Immunologic; Diphosphonates; Doxorubicin; Drug Screening Assays, Antitumor; Drug Synergism; Etoposide; Female; Genes, T-Cell Receptor delta; Genes, T-Cell Receptor gamma; Humans; Imidazoles; Interferon-gamma; Lovastatin; Lung Neoplasms; Male; Membrane Glycoproteins; Neoplasms; Perforin; Pore Forming Cytotoxic Proteins; Prostatic Neoplasms; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; Urinary Bladder Neoplasms; Vincristine; Zoledronic Acid

Recently, the scanning force microscope (SFM) has been widely used for direct monitoring of specific interactions between biologically active molecules. Such studies have employed the SFM liquid-cell setup, which allows measurements to be made in the native environment with force resolution down to a tenth of a picoNewton. In this study, the ligand-receptor strength of monoclonal anti-human prostatic acid phosphatase and prostatic acid phosphatase, representing an antigen-antibody system with a single type of interaction, was determined. Then, the interaction force occurring between concanavalin A and the carbohydrate component of the glycoproteins arylsulfatase A and carboxypeptidase Y was measured. High mannose-type glycans were sought on the human prostate carcinoma cell surface. Application of an analysis based on the Poisson distribution of the number of bonds formed in all these measured systems allowed the strength of the molecular interaction to be calculated. The values of the force acting between two single molecules were 530+/-25, 790+/-32, and 940+/-39 pN between prostatic acid phosphatase and monoclonal anti-human prostatic acid phosphatase, between concanavalin A and arylsulfatase A, and between concanavalin A and carboxypeptidase Y, respectively. The value calculated from data collected for the force between concanavalin A and mannose-containing ligands present on the surface of human prostate carcinoma cells was smaller, 116+/-17 pN. The different values of the binding force between concanavalin A and mannose-containing ligands were attributed to the structural changes of the carbohydrate components.

Topics: Binding Sites; Cell Adhesion; Cell Line, Tumor; Concanavalin A; Humans; Ligands; Male; Mannose; Micromanipulation; Microscopy, Atomic Force; Physical Stimulation; Prostatic Neoplasms; Protein Binding; Protein Conformation; Stress, Mechanical

Differences between human prostate carcinoma (PCA, five cases) and benign prostatic hyperplasia (BPH, five cases) in asparagine-linked (Asn) sugar-chain structure of prostatic acid phosphatase (PAP) were investigated using lectin affinity chromatography with concanavalin A (Con A) and wheat germ agglutinin (WGA). PAP activities were significantly decreased in PCA-derived PAP, while no significant differences between the two PAP preparations were observed in the enzymatic properties (Michaelis-Menten value, optimal pH, thermal stability, and inhibition study). In these PAP preparations, all activities were found only in the fractions which bound strongly to the Con A column and were undetectable in the Con A unbound fractions and in the fractions which bound weakly to the Con A column. The relative amounts of PAP which bound strongly to the Con A column but passed through the WGA column, were significantly greater in BPH-derived PAP than in PCA-derived PAP. In contrast, the relative amounts of PAP which bound strongly to the Con A column and bound to the WGA column, were significantly greater in PCA-derived PAP than in BPH-derived PAP. The findings suggest that Asn-linked sugar-chain structures are altered during oncogenesis in human prostate and also suggest that studies of qualitative differences of sugar-chain structures of PAP might lead to a useful diagnostic tool for PCA.

Topics: Acid Phosphatase; Adenocarcinoma; Aged; Chromatography, Affinity; Concanavalin A; Humans; Male; Middle Aged; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Wheat Germ Agglutinins

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

Topics: Antigens, Neoplasm; Concanavalin A; Diagnosis, Differential; Humans; Male; Mass Screening; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms; Serologic Tests

The enzymatic and immunological nature, and the sugar chain structure, of gamma-glutamyl transferase (GGT) purified from tissues of benign prostatic hypertrophy (BPH), prostatic carcinoma (PCa) and renal cell carcinoma (RCa), were compared with those of the normal prostate (NP) and kidney (NK). The specific activities of GGTs in NP, NK, BPH, PCa and RCa were 78.9, 22.5, 105, 92.5 and 52.5 mU/mg protein, respectively. The molecular masses of GGTs from BPH, PCa and RCa were 72 kDa, 78 and 108 kDa, and 79 and 105 kDa, respectively. The Michaelis constants (Km), optimum pHs and the inhibition of GGT activities by several chemical compounds, revealed that the GGT from BPH, PCa and RCa was similar to that of normal GGT. Immunologically, the IgG fraction against anti-human seminal plasma GGT fused to the all of the GGTs tested. The sugar chain heterogeneities of the various GGTs, detected by the serial-lectin affinity technique, differed from one another. The sugar chain of GGT from BPH resembled the sugar chain from NP. On the contrary, the sugar chains of GGTs from PCa and RCa were markedly different from those from normal tissues. In the GGT from PCa, multi-antennary complex type sugar chains were more increased than the enzyme of NP. In general, as previously reported, the sugar chains of GGTs from carcinomatous tissues of prostate and kidney had an increased content of bisecting GlcNAc (beta 1-->4) containing complex type sugar chains. Moreover, the reductions of the biantennary complex type sugar chain with fucose linkage and the hybrid type sugar chain were obvious in the GGT from carcinomatous tissues of the prostate and kidney.

Topics: Adult; Aged; Chromatography, Affinity; Concanavalin A; gamma-Glutamyltransferase; Humans; Hydrogen-Ion Concentration; Immunodiffusion; Kidney; Kidney Neoplasms; Kinetics; Lectins; Male; Middle Aged; Molecular Weight; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms

Topics: Antigens, Neoplasm; Concanavalin A; Diagnosis, Differential; Humans; Male; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms

The percentage of nonglycosylated prostate-specific antigen (PSA) was measured in the serum of 15 prostate cancer patients and 15 patients with benign hyperplasia of the prostate. The larger part of serum PSA in both groups was glycosylated, but while in carcinoma of the prostate the mean percentage of nonglycosylated PSA was 38.4 +/- 6.5, in benign prostate hyperplasia (BPH) only a mean of 14.2 +/- 4.3% of the PSA was nonglycosylated. These significantly higher results (p less than 0.001) suggest a different pattern of release of PSA from cancer cells and from hyperplastic or normal cells. Since in a part of the BPH we encounter elevations of PSA similar to the levels found in neoplasms, the degree of concanavalin A binding can provide an additional means in differentiating between benign and malignant lesions.

Topics: Antigens, Neoplasm; Concanavalin A; Glycosylation; Humans; Male; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms

The binding of different lectins (concanavalin A [Con A], triticum vulgaris [WGA], glycine maximum [SBA], dolichos bilflorus [DBA], ulex europaeus [UEA I], arachis hypogaea [PNA], and ricinus communis [RCA I]) to cells of normal prostate glands, hyperplastic glands and adenocarcinoma was studied. The Con A, WGA, DBA, PNA and RCA I bound to both normal and hyperplastic glands. The binding in the malignant glands differed from that of the benign conditions. The SBA, which was not bound by benign cells, was bound to the malignant glandular cells. Also, UEA I was bound to a part of the carcinoma cells. In addition, the binding pattern of Con A and WGA in the cells differed between the malignant and benign conditions. Based on the results of this study, it is suggested that lectin histochemical study might be useful in routine pathologic examination to detect malignant cells in cases which are doubtful with regard to malignancy by routine methods.

Topics: Adenocarcinoma; Biomarkers, Tumor; Concanavalin A; Fluorescein-5-isothiocyanate; Fluoresceins; Histocytochemistry; Humans; Lectins; Male; Peanut Agglutinin; Plant Lectins; Prostatic Hyperplasia; Prostatic Neoplasms; Soybean Proteins; Thiocyanates; Wheat Germ Agglutinins

We have utilized the technique of lectin-loading of SDS gels with iodinated concanavalin A and wheat germ agglutinin to identify glycoproteins in prostatic and seminal fluids as well as in prostate tissue fractions. The following subunits which bound both lectins were detected: (a) 50, 43 and 38 kDa subunits common to prostatic and seminal fluids, and an additional 55 kDa subunit which predominates only in prostatic fluid; (b) 78, 55, 50 and 43 kDa subunits in prostatic tissue cytosol and (c) 195, 170, 135, 116 and 95 kDa subunits present in the particulate fractions of prostatic tissue. Immunoblotting using specific rabbit antibodies revealed the 50 kDa band to be prostatic acid phosphatase and the 38 kDa band to be prostate-specific antigen. Interestingly, antibodies directed toward prostatic acid phosphatase were found to cross-react with the 43 kDa band. Fractionation on sucrose gradients showed that several of these particulate glycoproteins were associated with a vesicle fraction enriched in adenylate cyclase activity, implying that they are plasma membrane glycoproteins. Comparison of soluble and particulate fractions of normal and cancerous tissue homogenates was made by densitometric scanning of autoradiograms of lectin-loaded gels. Similar relative intensities of lectin-binding were obtained for corresponding proteins in normal and cancerous tissue fractions. Also, immunoblotting showed no differences in prostatic acid phosphatase or prostate-specific antigen between normal and cancerous soluble homogenate fractions. Our results suggest that major lectin-binding proteins are conserved in the transition from normal to cancerous tissue. These results may be useful in developing a multiple-marker profile of metastatic prostate cancer and for the design of imaging agents, such as monoclonal antibodies, to prominent soluble and particulate prostate glycoproteins.

Topics: Acid Phosphatase; Adenylyl Cyclases; Body Fluids; Cell Membrane; Concanavalin A; Cytosol; Electrophoresis, Polyacrylamide Gel; Glycoproteins; Humans; Immunoenzyme Techniques; Lectins; Male; Molecular Weight; Prostate; Prostatic Neoplasms; Semen; Wheat Germ Agglutinins

The responsiveness to diethylstilbestrol (DES) and estramustinephosphate (EMP) of human peripheral blood lymphocytes to T-cell mitogens has been investigated in vitro and in vivo. EMP demonstrated potent inhibition of both Con A- and PHA-induced lymphocyte proliferation in vitro, while it had no detectable effects when given to patients with cancer of the prostate. DES reduced the response to Con A in vitro, but had only marginal effects on PHA-induced mitogen response. In contrast, the response to Con A was unaltered, while the response to PHA was significantly diminished after DES therapy in patients with prostatic cancer. This effect, however, was only seen when high doses of DES not included in conventional regimen were given. The proliferative response to T-cell mitogens in patients with prostatic cancer was not affected by serum source in the assay, indicating the absence of humoral factors able to inhibit mitogen response in these patients.

Topics: Adult; Concanavalin A; Diethylstilbestrol; Estramustine; Humans; In Vitro Techniques; Lymphocyte Activation; Male; Middle Aged; Nitrogen Mustard Compounds; Phytohemagglutinins; Prostatic Neoplasms

Topics: Animals; Carrier Proteins; Castration; Cells, Cultured; Concanavalin A; Disease Models, Animal; Horseradish Peroxidase; Humans; Hydrolases; Lysosomes; Male; Neoplasm Proteins; Prostatic Neoplasms; Rats; Receptors, Retinoic Acid; Tretinoin

Concanavalin A (Con A)-inducible suppressor cell activity in peripheral blood lymphocytes (PBL) of urologic cancer patients and of appropriate controls with benign urologic disorders was measured concurrently. Although the proliferative responses to Con A of the cancer patients were significantly lower than those of controls, no difference in Con A-induced suppressor cell activity was demonstrated between cancer patients and controls when tested under a variety of conditions. Moreover, regression analysis revealed no correlation between the proliferative response to Con A and suppressor cell activity in either cancer patients or controls. The results indicated that Con A-inducible suppressor cell activity was unaltered in urologic cancer patients and suggested that suppressor cells of the type that can be activated by Con A were not involved in the general immunologic impairment frequently associated with urologic cancer.

Topics: Adenocarcinoma; Concanavalin A; Female; Humans; Kidney Calculi; Kidney Neoplasms; Lymphocyte Activation; Male; Middle Aged; Prostatic Hyperplasia; Prostatic Neoplasms; Prostatitis; T-Lymphocytes, Regulatory; Urethral Stricture; Urinary Bladder Neoplasms; Urinary Tract Infections; Urogenital Neoplasms

Topics: Aged; Androgen-Binding Protein; Carrier Proteins; Concanavalin A; Dihydrotestosterone; Humans; Kinetics; Leydig Cells; Male; Middle Aged; Prostatic Neoplasms; Seminiferous Tubules; Testis

The ultrastructural organization of the cells of the transplantable prostatic adenocarcinoma of the rat, R3327H, has been studied on a cytochemical level. The techniques used were those to localize lipids, peroxidase, and carbohydrate-like substances. This study has clearly shown that there are, in the tumor cells and not in normal prostatic epithelial cells, both lipid compartments with an intense peroxidase membrane. Within the lipid "droplet" we find it to be compartmentalized by conconavalan A-positive material. These observations suggest that this tumor cell may be a "degenerating" or old cell. It may well be that this cancer is a disease of dying cells--the degrading consequence of cell growth.

Topics: Adenocarcinoma; Animals; Carbohydrates; Cell Survival; Concanavalin A; Histocytochemistry; Intracellular Membranes; Lipids; Male; Neoplasms, Experimental; Peroxidases; Prostatic Neoplasms; Rats

We have recently described the presence of a guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] inhibitor (GCI) in an aqueous extract of the balsam pear (Momordica charantia abbreviata). Because the guanylate cyclase-cyclic GMP system is though to be involved in cell growth, DNA and RNA synthesis, and possible malignant transformation, we examined the effect of the aqueous extract containing GCI on an undifferentiated adenocarcinoma of the rat prostate and concanavalin-A-stimulated [3H]thymidine incorporation into cultured splenic lymphocytes, a process thought to be mediated by cyclic GMP. The results demonstrate that the extract of the balsam pear blocks both the growth of the rat prostatic adencarcinoma in vitro and [3H]thymidine incorporation into DNA. DNA histograms from flow cytometry indicated that the extract containing GCI inhibited in the G2 + M phase of the cell cycle, a presumed locus of cyclic GMP effects. In addition, guanylate cyclase activity was significantly greater in the tumor than normal prostate tissue and was decreased by the extract containing GCI. Cyclic GMP levels in the tumor in culture wer also decreased by addition of the extract. It remains to be determined whether or not the anti-tumor agent and GCI are the same substance.

Topics: Adenocarcinoma; Animals; Cell Cycle; Concanavalin A; Cyclic GMP; Guanylate Cyclase; In Vitro Techniques; Male; Neoplasms, Experimental; Plant Extracts; Prostatic Neoplasms; Thymidine

Concanavalin A and wheat germ agglutinin, lectins that interact with serum glycoprotein in a manner similar to the antigen--antibody reaction, were used as "antibodies" in a single radial immunodiffusion technique to test a coded serum panel (from the National Cancer Institute, Bethesda, Md., and the Mayo Clinic, Rochester, Minn.) containing a) 99 serum samples from patients with different types of malignant neoplasms of the gastrointestinal tract, prostate gland, and lung, b) 50 samples from patients with benign diseases of the same organs as those affected in the cancer patients, and c) 50 samples from apparently healthy smokers. The resulting precipitation rings were not correlated to serum protein concentration, and the differences (demonstrated by Student's t-test and with a generalization of the one-sided two-sample Kolmogorov-Smirnov statistic for evaluating diagnostic tests) established that serum glycoproteins are glycosylated differently in cancer patients than in people without cancer.

Topics: Blood Proteins; Concanavalin A; Female; Gastrointestinal Neoplasms; Glycoproteins; Humans; Immunodiffusion; Lectins; Lung Neoplasms; Male; Neoplasm Proteins; Neoplasms; Prostatic Neoplasms