dinitrobenzenes and Carcinoma--Hepatocellular

The erythropoietin-producing hepatocellular carcinoma receptor B4 is a receptor tyrosine kinase whose expression is preserved in various malignancies, including colon, gastric, and breast carcinoma. Hepatocellular carcinoma receptor B4 presence in tumor cells and involvement in cancer suppression makes it a potential therapeutic target for activating compounds. Moreover, modulators of its activity also have a strong potential to be used in diagnosis and therapy monitoring. We used virtual ligand screening to identify novel hepatocellular carcinoma receptor B4 kinase modulators for experimental testing. Three independent assay platforms confirmed that dinitrophenyl-L-arginine is likely to affect the kinase activity of hepatocellular carcinoma receptor B4. An enzyme-coupled spectrophotometric assay has been used to examine this possibility and may prove to be useful for assessing other novel kinase modulator candidates. Overall, our observations suggest that dinitrophenyl-L-arginine has an activating effect on hepatocellular carcinoma receptor B4 and, therefore, more efficient derivatives may have therapeutic effects in tumors where hepatocellular carcinoma receptor B4 exhibits antimalignant properties. The hepatocellular carcinoma receptor B4-activating effect is discussed with respect to previously described mechanisms, using predicted and experimental structures for docked ligands. As a novel kinase activity modulator, dinitrophenyl-L-arginine may provide new insights into molecular mechanisms by which kinases are activated or regulated, and may serve as a lead compound for the generation of novel hepatocellular carcinoma receptor B4-activating therapeutic compounds.

Topics: Antineoplastic Agents; Arginine; Carcinoma, Hepatocellular; Dinitrobenzenes; Humans; Liver Neoplasms; Neoplasm Proteins; Receptor, EphB4

Dinitrotoluenes (DNTs) are byproducts of the explosive trinitrotoluene (TNT), and exist as a mixture of 2 to 6 isomers, with 2,4-DNT and 2,6-DNT being the most significant. The main route of human exposure at ammunition facilities is inhalation. The primary targets of DNTs toxicity are the hematopoietic system, cardiovascular system, nervous system and reproductive system. In factory workers, exposure to DNTs has been linked to many adverse health effects, including: cyanosis, vertigo, headache, metallic taste, dyspnea, weakness and lassitude, loss of appetite, nausea, and vomiting. Other symptoms including pain or parasthesia in extremities, abdominal discomfort, tremors, paralysis, chest pain, and unconsciousness have been documented. An association between DNTs exposure and increased risk of hepatocellular carcinomas and subcutaneous tumors in rats, as well as renal tumors in mice, has been established. This research was therefore designed targeting the liver to assess the cellular and molecular responses of human liver carcinoma cells following exposure to 2,4-DNT and 2,6-DNT. Cytotoxicity was evaluated using the MTT assay. Upon 48 hrs of exposure, LC50 values of 245 +/- 14.724 microg/mL, and 300 +/- 5.92 microg/mL were recorded for 2,6-DNT and 2,4-DNT respectively, indicating that both DNTs are moderately toxic, and 2,6-DNT is slightly more toxic to HepG2 cells than 2,4-DNT. A dose response relationship was recorded with respect to the cytotoxicity of both DNTs. Western blot analysis resulted in a significant expression (p<0.05) of the 70-kDa heat shock protein in 2,6-DNT-treated cells compared to the control cells and at the 200 microg/mL dose for 2,4-DNT. A statistically significant expression in c-fos was also observed at the 200 and 250 microg/mL treatment level for 2,4- and 2,6-DNT, respectively. However, no statistically significant expression of this protooncogene-related protein was observed at the doses of 0, 100, or 300 microg/mL or within the dose range of 0-200 microg/mL for 2,6-DNT. The 45-kDa growth arrest and damage protein was significantly expressed at the dose range of 200 - 250 microg/mL for 2,6-DNT and at the dose range of 200 - 400 microg/mL for 2,4-DNT. Expression of 153-kDa growth arrest and DNA damage protein was significant at the 100, 200, and 250 microg/mL doses for 2,6-DNT and at the 200 microg/mL dose for 2,4-DNT. Overall, these results indicate the potential of DNTs to induce cytotoxic, proteotoxic (HSP70), and genotoxic

Topics: Carcinoma, Hepatocellular; Cell Cycle Proteins; Cell Line, Tumor; Cell Survival; Dinitrobenzenes; HSP70 Heat-Shock Proteins; Humans; Nuclear Proteins; Proto-Oncogene Proteins c-fos; Transcription Factor CHOP

The nitro musk compounds musk xylene (1-tert.-butyl-3,5-dimethyl-2,4,6-trinitrobenzene), musk ketone (4-tert.-butyl-3,5-dinitro-2,6-dimethylacetophenone), musk ambrette (1-tert.-butyl-4-methyl-6-methoxy-3,5-dinitrobenzene), musk moskene (1,1,3,3,5-pentamethyl-4,6-dinitroindane) and musk tibetene (1-tert.-butyl-3,4,5-trimethyl-2,6-dinitrobenzene) were tested for their genotoxic activity in the micronucleus test (MN) with human lymphocytes in vitro and the human hepatoma cell line Hep G2. Compound concentrations were employed up to cytotoxic doses. Musk xylene, musk ketone, musk ambrette, musk moskene and musk tibetene revealed no genotoxicity in the micronucleus test with human lymphocytes and with the human hepatoma cell line Hep G2.

Topics: Adult; Carcinoma, Hepatocellular; Cells, Cultured; Chi-Square Distribution; Dinitrobenzenes; Humans; Indans; Liver Neoplasms; Lymphocytes; Micronucleus Tests; Mutagens; Perfume; Structure-Activity Relationship; Tumor Cells, Cultured; Water Pollutants, Chemical; Xylenes

Conditions have been established for H4IIE rat hepatoma cell cultures in which effects of cytochrome P-450 induction on the metabolism of a munitions wastestream pollutant can be studied. Under these conditions, the polychlorinated hydrocarbon 2,3,4,7,8-pentachlorodibenzfuran (PCDBF) induced cytochrome P-450 (1A1) aryl hydrocarbon hydroxylase (AHH) activity over a wide range of concentrations without significant cytotoxic effects. The munition pollutant 2,4-dinitrotoluene (2,4-DNT) did not induce AHH activity itself, but its metabolism was considerably altered when applied to PCDBF induced cultures. Production of amino nitrotoluene isomers was greatly enhanced in induced cultures as compared to uninduced controls, as was the conversion of radiolabeled 2,4-DNT to relatively more polar metabolites. To some extent, the results with H4IIE cells parallel those reported for animals exposed to 2,4-DNT after induction of cytochrome P-450 AHH activity. The preliminary findings suggest that with further development and validation, H4IIE cultures could be of use in characterizing metabolites that result from exposure to chemical mixtures involving a P-450 (1A1) inducer.

Topics: Animals; Aryl Hydrocarbon Hydroxylases; Benzofurans; Carcinoma, Hepatocellular; Cell Survival; Cytochrome P-450 Enzyme System; Dinitrobenzenes; Enzyme Induction; Liver Neoplasms; Luminescent Measurements; Mutagenicity Tests; Oxidation-Reduction; Rats; Spectrophotometry, Ultraviolet; Tumor Cells, Cultured

An analysis of the mortality experience of workers exposed to dinitrotoluene (DNT) was conducted to test the hypothesis that DNT exposure is associated with an increased risk of cancers of the liver and biliary tract. A total of 4,989 workers exposed to DNT and 7,436 unexposed workers who had worked for at least 5 months at the study facility between January 1, 1949 and January 21, 1980, were included in this investigation. Workers were considered exposed if they had worked at least 1 day on a job with probable exposure to DNT. The vital status as of December 31, 1982, was successfully ascertained for approximately 97% of these workers. Standardized mortality ratios (SMRs) were estimated based upon comparisons with the US population using a modified life-table program. In addition, standardized rate ratios (SRRs) were computed based upon direct comparisons between the DNT and the internal unexposed cohort. An excess of hepatobiliary cancer was observed among workers exposed to DNT in this study. The rate ratio for hepatobiliary cancer was 2.67 (six cases observed) based upon comparison with the US population (SMR = 2.67, 95% CI = 0.98, 5.83), and 3.88 based upon comparison using the internal unexposed referent group (SRR = 3.88, 95% CI = 1.04, 14.41). This study failed to demonstrate an exposure-response relationship between duration of DNT exposure and hepatobiliary cancer mortality. Our study was limited by the small number of workers with long duration of exposure to DNT, and by the lack of quantitative information on exposure to DNT and other chemicals.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Air Pollutants, Occupational; Bile Duct Neoplasms; Carcinogens; Carcinoma, Hepatocellular; Cause of Death; Cohort Studies; Dinitrobenzenes; Humans; Life Tables; Liver Neoplasms; Male; Middle Aged; Neoplasms; Occupational Diseases; Occupational Exposure; Retrospective Studies; Survival Analysis

Hydrolysis of extramitochondrial ATP by coupled Zajdela hepatoma mitochondria is not stimulated by uncouplers of oxidative phosphorylation. The results of the present study show that the hydrolysis of intramitochondrial ATP in these mitochondria is stimulated by DNP and CCCP. It is proposed that the uncoupler insensitivity of ATPase in coupled Zajdela hepatoma mitochondria with exogenous ATP as a substrate result from an altered functional relationship between ATPase and ADP, ATP translocase.

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Animals; Carbonyl Cyanide m-Chlorophenyl Hydrazone; Carcinoma, Hepatocellular; Dinitrobenzenes; Hydrolysis; Liver Neoplasms; Mitochondria, Liver; Rats; Uncoupling Agents