phosphorus-radioisotopes and Urinary-Bladder-Neoplasms

A multiple biomarker approach is required to integrate for metabolism, temporal response and exposure-response kinetics, biological relevance, and positive predictive value. Carcinogen DNA adduct analysis can be used in animal and in vitro studies to detect absorption permutations caused by mixture interactions, and to control metabolic variation when specific CYP450 genes (1A1 or 1A2) are knocked out. These enzymes are not critical to the metabolic activation of model Polycyclic Aromatic Compounds (PAC) and aromatic amines, respectively, as suggested by in vitro analysis. Several human studies have been carried out where multiple biomarkers have been measured. In a study of benzidine workers, the similarities in elimination kinetics between urinary metabolites and mutagenicity is likely responsible for a better correlation between these markers than to BZ-DNA adducts in exfoliated cells. In a study of rubber workers, the relationship between specific departments, urinary 1 HP and DNA adducts in exfoliated cells coincided with the historical urinary bladder cancer risk in these departments; the same relationship did not hold for urinary mutagenicity. In a study of automotive mechanics, biomarkers were used to monitor the effectiveness of exposure interventions. These data reinforce the notion that carcinogen biomarkers are useful to monitor exposure, but that a complementary approaches involving effect and perhaps susceptibility biomarkers is necessary to obtain the necessary information.

Topics: Animals; Automobiles; Benzidines; Biomarkers, Tumor; Carcinogens; DNA; DNA Adducts; Environmental Monitoring; Epidemiological Monitoring; Humans; In Vitro Techniques; Industry; Mutagenicity Tests; Occupational Exposure; Phosphorus Radioisotopes; Rubber; Urinary Bladder Neoplasms; Urine

Cigarette smoking is an established cause of bladder cancer. The direct relationship between smoking-induced DNA adducts in bladder cells and cancer risk at that site has, however, been poorly assessed. We therefore investigated the relationship between bladder cancer risk and levels of DNA adducts measured in normal bladder biopsies by 32P-post-labeling in a hospital-based case-control study of 59 bladder cancer patients and 45 controls submitted to surgery for prostatic hyperplasia or urinary incontinence. An approximately 2-fold risk for bladder cancer was found in individuals with an adduct level >14.8 (median among controls) compared with those with an adduct level < or =14.8 (OR = 1.9, 95% CI 0.8-4.3, P = 0.13). A dose-response relationship was also suggested (trend test, P = 0.13): compared with adduct levels below 13.5, the OR for bladder cancer was 1.7 (95% CI 0.6-4.6) for adduct levels between 13.5 and 18.5 and 2.2 (95% CI 0.8-6.1) for adduct levels >18.5. These findings provide some evidence that DNA adducts in bladder tissue might predict smoking-induced bladder cancer. Larger studies are still warranted to confirm these results.

Topics: Aged; Biopsy; Case-Control Studies; DNA Adducts; Female; Humans; Leukocytes; Male; Phosphorus Radioisotopes; Predictive Value of Tests; Reference Values; Risk Factors; Smoking; Urinary Bladder; Urinary Bladder Neoplasms

DNA adducts formed in human uroepithelial cells (HUC) following exposure to N-hydroxy-4-aminobiphenyl (N-OH-ABP), the proximate metabolite of the human bladder carcinogen 4-aminobiphenyl (ABP), were analyzed by the (32)P-postlabeling method. Two adducts detected by (32)P-postlabeling were previously identified as the 3',5'-bisphospho derivatives of N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG-C8-ABP) and N-(deoxyadenosin-8-yl)-4-aminobiphenyl (dA-C8-ABP) (Frederickson S et al. [1992] Carcinogenesis 13: 955-961; Hatcher and Swaminathan [1995b] Carcinogenesis 16: 295-301). In contrast to the dG-C8-ABP adduct, which was 3'-dephosphorylated by nuclease P1, dA-C8-ABP was resistant to nuclease P1, thus providing an enrichment step before postlabeling. Autoradiography of the two-dimensional thin-layer chromatogram of the postlabeled products obtained following nuclease P1 digestion revealed several minor adducts, one of which has been identified in the present study. Postlabeling analyses following nuclease P1 digestion of the products obtained from the reaction of N-acetoxy-4-aminobiphenyl with deoxyguanosine-3'-monophosphate (dGp) demonstrated the presence of this minor adduct. The 3'-monophosphate derivative of the adduct was subsequently chromatographically purified and subjected to spectroscopic analyses. Based on proton NMR and mass spectroscopic analyses of the synthetic product, the chemical structure of the adduct has been identified as N-(deoxyguanosin-N(2)-yl)-4-azobiphenyl (dG-N==N-ABP). (32)P-Postlabeling analysis of the nuclease P1-enriched DNA hydrolysate of HUCs treated with N-OH-ABP or N-hydroxy-4-acetylaminobiphenyl (N-OH-AABP) showed the presence of the dG-N==N-ABP adduct. It was also detected in calf thymus DNA incubated with HUC cytosol and N-OH-ABP in the presence of acetyl-CoA, or incubated with HUC microsomes and N-OH-AABP. These results demonstrate that in the target cells for ABP carcinogenesis in vivo, N-OH-ABP and N-OH-AABP are bioactivated by acyltransferases to reactive arylnitrenium ions that covalently interact at the N2 position of deoxyguanosine in DNA.

Topics: Aminobiphenyl Compounds; Azo Compounds; Biphenyl Compounds; Carcinogens, Environmental; Cells, Cultured; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Deoxyribonucleotides; DNA; DNA Adducts; Humans; Magnetic Resonance Spectroscopy; Phosphorus Radioisotopes; Urinary Bladder Neoplasms; Urothelium

Examination of microsatellites is frequent in the diagnosis of cancer. Microsatellites are repeat DNA sequences scattered throughout the human genome. These repeat regions are very frequent and highly polymorphic elements. In this study we focus on dinucleotide repeats. We compared three different methods for the detection of microsatellites: use of the ABI Prism 377 fluorescence sequencer, autoradiography and silver-stained gels. DNA was extracted from various clinical samples and amplified by different polymerase chain reaction (PCR) protocols. DNA from normal and tumor tissues was analysed using each method. The fluorescence method was more sensitive than the two other methods; however, this technology is very expensive. It seems possible, when examining microsatellites on a low budget, to avoid radioactivity by using silver-stained gels as an alternative. In conclusion, we observed identical results when comparing autoradiography with the fluorescence technique. However, we observed variability in the results when interpreting a single locus comparing silver staining with autoradiography and the fluorescence technique. Classification of the tumors based on several microsatellite loci was always identical.

Topics: Autoradiography; Colonic Neoplasms; DNA Primers; DNA, Neoplasm; DNA, Satellite; Fluorescent Dyes; Genetic Markers; Humans; Phosphorus Radioisotopes; Polymerase Chain Reaction; Repetitive Sequences, Nucleic Acid; Sequence Analysis, DNA; Silver Staining; Urinary Bladder Neoplasms

Recent studies have demonstrated the presence of DNA adducts from 4-aminobiphenyl (4-ABP) in the bladder cells of humans; however, the correlation between the concentration of these adducts and the tumorigenic response is not clear. To help elucidate this relationship, we have investigated DNA adduct formation in experimental animals continuously administered 4-ABP. Male and female BALB/c mice were treated for 28 days with 4-ABP. hydrochloride in their drinking water. DNA adducts in target tissues (liver of females and bladder of males) were identified and quantified by 32P-postlabeling analyses and radioimmunoassays. These results were compared to previously reported tumor incidences obtained from the lifetime administration of 4-ABP hydrochloride. The major adduct observed in both tissues was N-(deoxyguanosin-8-yl)-4-ABP. In the bladders of both sexes and the livers of female mice, adduct levels increased with dose at low doses, but saturation was observed at high doses. In the livers of males, the adduct levels were linearly correlated with dose throughout the entire dose range. A comparison between DNA adducts and tumorigenesis indicated a linear correlation between adduct levels and the incidence of liver tumors in female mice. In the bladders of male mice, however, the relationship was markedly nonlinear. These data suggest that adduct formation alone is insufficient for tumorigenesis in the bladder and that other factors such as cell proliferation are necessary for tumor production.

Topics: Aminobiphenyl Compounds; Animals; DNA; DNA Adducts; Female; Liver Neoplasms, Experimental; Male; Mice; Mice, Inbred BALB C; Phosphorus Radioisotopes; Radioimmunoassay; Urinary Bladder Neoplasms

The effects of local irradiation and intraperitoneal injection of cisplatinum (CDDP) and VP-16 were examined in the sequential 31P magnetic resonance spectroscopy (MRS) in testicular cancer (TC-1) and bladder tumor (BT-8) of human origin, serially transplanted in nude mice. In the early phase of tumor growth, high-energy phosphate metabolites such as phosphocreatinine (PCr), adenosine triphosphate (ATP) and phosphomonoester (PME) were detected in both grafted tumors. However, the relative value of inorganic phosphate (Pi) to PCr increased with the growth of the tumor. Irradiation had the most pronounced effect to inhibit growth, followed by CDDP in both strains. However, growth inhibition was not observed in the VP-16 group. The effect of irradiation on the tumor histology was severely expressed in the nucleus and cytoplasm on the 4th to 7th day. The high PCr/Pi ratio during 2 to 14 days after irradiation suggested reoxygenation in the tumors with a high hypoxic cell fraction. In the CDDP and VP-16 groups, without histological change, the changes of PCr and Pi were milder than that in the irradiation group. Thus the spectroscopic analysis is presumably expected to give us an earlier and more accurate information on the tumor than the conventional parameters.

Topics: Adenosine Triphosphate; Animals; Antineoplastic Combined Chemotherapy Protocols; Humans; Magnetic Resonance Spectroscopy; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Phosphocreatine; Phosphorus Radioisotopes; Testicular Neoplasms; Urinary Bladder Neoplasms

Ochratoxin A (OTA), a natural contaminant of mouldy food and feed, is suspected of being one of the etiological agents responsible for Balkan endemic nephropathy (BEN) and the associated urinary tract tumours. We have previously shown that ochratoxin A is genotoxic as expressed by DNA single-strand breaks. DNA-OTA adducts have been detected in various mouse organs after ochratoxin A treatment. Tumorous tissues from three kidneys and five bladders of Bulgarian patients undergoing surgery for cancer and from three non-malignant kidneys collected from French subjects were analysed for DNA adducts. Several adducts with the same RF values as those obtained from mouse kidney after treatment with OTA (one major and some minor adducts) were detected, mainly in kidney but also in bladder tissues from Bulgaria. No adducts were detected in French kidney tissues. These results provide new evidence of the possible role of OTA in the development of tumours of the urinary tract in Bulgaria.

Topics: Animals; Balkan Nephropathy; Bulgaria; DNA; DNA Damage; DNA, Neoplasm; Humans; Kidney; Kidney Neoplasms; Mice; Ochratoxins; Phosphorus Radioisotopes; Urinary Bladder Neoplasms

4-Aminobiphenyl (4-ABP) is a human and mouse bladder carcinogen. Epidemiological studies have shown that individuals with a slow acetylator phenotype, especially those exposed to high levels of carcinogenic aromatic amines, show an increased susceptibility to bladder cancer. In order to determine if a slow acetylator phenotype results in increased DNA damage, congenic mouse strains C57BL/6J and B6.A-Nat(s), which differ genetically at the acetyltransferase (EC 2.3.1.5) locus as homozygous rapid (Natr/Natr) and homozygous slow (Nat(s)/Nat(s)) acetylators respectively, were continuously administered 4-ABP.HCl (55-300 p.p.m.) in their drinking water for 28 days. The levels of covalently bound N-(deoxyguanosin-8-yl)-4-ABP-DNA adducts, which are believed to be critical for the initiation of tumors, were quantitated in the liver and bladder by 32P-postlabeling analysis. The levels of the hepatic DNA adduct increased with dose in both sexes, but were independent of the mouse acetylator genotype. At comparable doses, however, the levels of DNA adducts were 2-fold higher in the liver of the female as compared to the male animals. The DNA adducts also increased with dose in bladder of the male mice, but in contrast to the liver, the adduct levels were approximately 2-fold lower in the bladder DNA of the female mice. Also in contrast to the liver, the levels of bladder DNA adducts were significantly higher (P < or = 0.03) in the phenotypic rapid acetylator females compared to the slow acetylators at both 75 and 150 p.p.m. doses; the median levels of adducts were 10-20% higher in the phenotypic slow acetylator male bladders compared to their rapid acetylator counterparts. The results of these studies are consistent with the increased carcinogenicity of 4-ABP to the liver of female mice and the bladder of male mice. They further suggest that factors other than acetylator phenotype limit the extent of DNA adduct formation from 4-ABP in these mice.

Topics: Acetylation; Aminobiphenyl Compounds; Animals; Autoradiography; Deoxyguanosine; DNA; DNA Damage; Female; Genotype; Isotope Labeling; Liver; Male; Mice; Mice, Inbred C57BL; Phenotype; Phosphorus Radioisotopes; Time Factors; Urinary Bladder; Urinary Bladder Neoplasms

Phosphoproteins of control and IFN-beta treated human bladder carcinoma cells (RT4) were labelled in vitro with [32P]-ATP and analyzed by polyacrylamide gel electrophoresis. Cells treated with antiproliferative doses of IFN had reduced levels of phosphorylated 60 Kd and 40 Kd proteins. IFN also induced within 24 hours the modification of a low molecular weight phosphoprotein doublet in the 22-24,000 molecular weight range. The ability to phosphorylate high molecular weight proteins by the in vitro procedures was generally depressed by IFN treatment. There was a dramatic shift in the phosphorylation of alkali stable phosphoamino acids associated with proteins in the 43-50,000 molecular weight range in IFN-treated cells. Preliminary studies indicate that at least some of the IFN-induced modifications of cellular phosphoproteins may result from transcriptional control of specific oncogenes.

Topics: Adenosine Triphosphate; Cell Cycle; Cell Line; Humans; Interferon-gamma; Kinetics; Molecular Weight; Phosphoproteins; Phosphorus Radioisotopes; Urinary Bladder Neoplasms

The prevention of recurrences of bladder cancer was attemped in 48 patients by means of the combined intravesical instillation of thio-tepa and urokinase and in 28 patients through the instillation of thio-tepa alone. The recurrence rates of both therapies for the postoperative 18 months were 7.9 and 32.6 per cent, respectively, indicating a significant drop in the recurrence rate in the group subjected to the combined therapy. No significant difference was found between the 2 instillation groups in terms of the blood transmission of 32-P thio-tepa. Serious leukopenia was found in 2 of the 48 patients receiving the combined instillation therapy but we concluded that this was not attributable to the use of urokinase.

Topics: Adolescent; Adult; Aged; Carcinoma, Transitional Cell; Cystitis; Drug Therapy, Combination; Endopeptidases; Female; Hemorrhage; Humans; Leukopenia; Male; Middle Aged; Neoplasm Recurrence, Local; Phosphorus Radioisotopes; Thiotepa; Urinary Bladder Diseases; Urinary Bladder Neoplasms; Urokinase-Type Plasminogen Activator

Bladders of normal rats were used to evaluate the absorption of thio-tepa after intravesical instillation. Thio-tepa labeled with 32P (radioactive phosphorus) was utilized to measure drug concentrations in bladder wall, liver, and bone marrow. Thio-tepa was rapidly absorbed into the general circulation, and significant amounts of isotope were found. Thio-tepa absorption patterns in the normal rat may serve as a guide in adjusting drug dosages when thio-tepa is used intravesically in human beings for the treatment of superficial, papillary bladder tumors.

Topics: Absorption; Administration, Intravesical; Animals; Female; Phosphorus Radioisotopes; Rats; Rats, Inbred F344; Thiotepa; Urinary Bladder; Urinary Bladder Neoplasms

Topics: Cobalt; Cobalt Radioisotopes; Humans; Nylons; Phosphorus; Phosphorus Radioisotopes; Sutures; Urinary Bladder Neoplasms

Topics: Chromium; Chromium Compounds; Humans; Phosphates; Phosphorus; Phosphorus Radioisotopes; Phosphorus, Dietary; Urinary Bladder Neoplasms

Topics: Chromium; Chromium Compounds; Humans; Phosphates; Phosphorus; Phosphorus Radioisotopes; Urinary Bladder Neoplasms; Urologic Neoplasms

Topics: Chromium; Chromium Compounds; Humans; Phosphates; Phosphorus; Phosphorus Radioisotopes; Phosphorus, Dietary; Urinary Bladder Neoplasms

Topics: Chromium; Chromium Compounds; Neoplasms; Phosphates; Phosphorus; Phosphorus Radioisotopes; Urinary Bladder Neoplasms

Topics: Gold Radioisotopes; Humans; Male; Phosphorus; Phosphorus Radioisotopes; Prostatic Neoplasms; Radioisotopes; Urinary Bladder Neoplasms; Urology