Animals ; Apoptosis ; drug effects ; Benzopyrenes ; administration & dosage ; toxicity ; Hippocampus ; drug effects ; pathology ; Injections, Intraventricular ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo clone differentially expressed cDNA sequences involved in malignant transformation induced by benzo(a)pyrene metabolite dihydroxyepoxy benzo pyrene (BPDE).

METHODThe malignant transformation of human bronchial epithelial cell line 16HBE induced by BPDE in vitro was used as a model for comparing gene expression between the transformed cells and controls. cDNA representational difference analysis (cDNA-RDA) was performed to isolate differentially expressed cDNA fragment in transformed cells. The cDNA fragments were ligated to pGEM-T vector and transformed into JM109 bacteria. The plasmid DNA were sequenced and compared with data in GenBank by BLASTN.

RESULTSFive cDNA sequences were found to be novel ones and were registered in dbest database, which assigned accession numbers in GenBank are BG354691, BG354692, BG354693, BG354694 and BG354695, respectively. Eight of the remaining cDNA sequences showed sequence homology to those previously reported such as ribosomal protein S23, MLN137, ACTN4, transforming growth factor and G protein gene.

CONCLUSIONSThese 13 genes may be involved in BPDE-induced malignant transformation, but their biological characteristics and functions are left to further studies.

Benzopyrenes ; metabolism ; pharmacology ; Carcinogens ; pharmacology ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; Cells, Cultured ; Cloning, Molecular ; DNA, Complementary ; analysis ; drug effects ; DNA, Neoplasm ; analysis ; Gene Expression ; drug effects ; Humans

OBJECTIVETo study the modulatory effect of distillate of Ocimum sanctum (traditionally known as Tulsi) leaf extract (DTLE) on genotoxicants.

METHODSIn the present investigation, we studied the antigenotoxic and anticlastogenic effect of distillate of Tulsi leaf extract on (i) human polymorphonuclear leukocytes by evaluating the DNA strand break without metabolic activation against mitomycin C (MMC) and hexavalent chromium (Cr+6) and (ii) human peripheral lymphocytes (in vitro) with or without metabolic activation against mitomycin C (MMC), hexavalent chromium (Cr+6) and B[a]P by evaluating chromosomal aberration (CA) and micronucleus assay (MN). Three different doses of DTLE, 50 microL/mL, 100 microL/mL, and 200 microL/mL were selected on the basis of cytotoxicity assay and used for studying DNA strand break, chromosomal aberration and micronucleus emergence. The following positive controls were used for inducing genotoxicity and clastogenicity: MMC (0.29 micromol/L) for DNA strand break, chromosomal aberration and 0.51 micromol/L for micronucleus assay; Potassium dichromate (Cr+6) 600 micromol/L for DNA strand break and 5 micromol/L for chromosomal aberration and micronucleus assay; Benzo[a]pyrene (30 micromol/L) for chromosomal aberration and 40 micromol/L for micronucleus assay. The active ingredients present in the distillate of Tulsi leaf extract were identified by HPLC and LC-MS.

RESULTSMitomycin C (MMC) and hexavalent chromium (Cr+6) induced statistically significant DNA strand break of respectively 69% and 71% (P<0.001) as revealed by fluorometric analysis of DNA unwinding. Furthermore, the damage could be protected with DTLE (50 microL/mL, 100 microL/mL, and 200 microL/mL) on simultaneous treatment. Chromosomal aberration and micronucleus formation induced by MMC, Cr+6 and B[a]P were significantly protected (P<0.001) by DTLE with and without metabolic activation.

CONCLUSIONDistillate of Tulsi leaf extract possesses antioxidants contributed mainly by eugenol, luteolin and apigenin as identified by LC-MS. These active ingredients may have the protective effect against genotoxicants.

Adult ; Benzopyrenes ; toxicity ; Cell Survival ; drug effects ; Chromium ; toxicity ; Chromosome Aberrations ; drug effects ; DNA ; metabolism ; DNA Damage ; drug effects ; Humans ; Lymphocytes ; drug effects ; Mass Spectrometry ; Mitomycin ; toxicity ; Mutagens ; toxicity ; Ocimum ; chemistry ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry

No abstract available.
Adenoma* ; Animals ; Benzo(a)pyrene* ; Ganoderma* ; Mice*

To evaluate the difference of concentration and mutagenicity of organic pollutants between residential and traffic area of Seoul, air samples were collected in Bulkwang (residential) and Shinchon (traffic) area. Samples were analyzed to measure the concentration of extractable organic matters (EOM) and their subfractions and mutagenicites were tested using Salmonella typhimurium TA 98. The concentrations of polycyclic aromatic hydrocarbons (PAHs) were also measured by gas-chromatography and compared between two areas. The results were as follows ; 1. While the concentration of total suspended particulate (TSP) in residental area was below the environmental standard in annual average, the concentration in traffic area was above the standard and was up to its maximum 256 microgram/m3 in November. The difference of TSP concentrations in both areas of each month was statistically significant (P<0.05). 2. The concentration of fine particle in traffic area was significantly higher compare to that in residential area and showed statistically significant monthly difference in both areas (P<0.05). The proportion of concentration of fine particle to TSP was 55-68%. 3. Mean concentrations of EOM in residential and traffic areas were 4.3 microgram/m3 and 5.3 microgram/m3 respectively. The proportion of amount of EOM from fine particle to EOM from TSP was 70-88%. 4. While the percentage of polar neutral organic compounds (POCN) of fine particle in Bulkwang's sample was higher compare to Shinchon's sample, the percentage of aliphatic compounds of fine particle in Shinchon's sample was higher compare to Bulkwang's sample. The percentages of PAH fraction were as low as 6-10% in both areas. 5. The mutagenic activity of unit concentration of organic matters extracted from fine particle was higher compare to that of coarse particle and was increased when metabolically activated with S9. Mutagenicities with metabolic activation calculated by unit air volume were significantly different between residential and traffic area, 17 revertants/m3 and 22 revertants/m3 respectively. 6. The concentrations of benzo(a)pyrene in fine particle of traffic and residential areas were 3.10 microgram/m3 and 2.02 microgram/m3 respectively. Sixteen PAHs were higher in samples of traffic area compare to residential area and also concentrations of PAHs in fine particle were higher compare to coarse particle.
Benzo(a)pyrene ; Biotransformation ; Polycyclic Hydrocarbons, Aromatic ; Salmonella typhimurium ; Seoul*

Metabolomics methods were applied in the study of the toxicity of environmental pollutants. It has been shown that exposure to heavy metals such as mercury, arsenic, cadmium, chromium and lead could cause significant changes in energy, lipids, nucleic acids and amino acids in mammalian cells. After exposure to benzo(a)pyrene [B(a)P], the glands of Pinctada pumila could produce various changes, such as energy metabolic disorder, cell damage, signal transduction disorder, oxidative stress and osmotic disorder. Persistent organic compounds polychlorinated biphenyls (PCBs) could exert toxic effects on Zebrafish embryos through affecting amino acid metabolism, DNA and protein methylation and biosynthesis. After exposure to endocrine disruptors, such as nonylphenol, octylenediester phthalate and bisphenol propane, goldfish showed energy, lipid and nucleic acid metabolic disorders.
Animals ; Benzo(a)pyrene ; Environmental Pollutants ; Metabolomics ; Oxidative Stress

OBJECTIVETo explore the effect of miR-542-3p in malignant transformation of human bronchial epithelial cells (16HBE) induced by anti-benzo(a)pyrene-7,8-diol-9,10-epoxide (anti-BPDE).

METHODSThe relative expression level of mature miR-542-3p in transformed cells (16HBE-T) and untransformed control cells (16HBE-N) was measured by real-time quantitative polymerase chain reaction (qRT-PCR). miRNA mimic was transiently transfected into 16HBE-T to change the expression level of miR-542-3p, and then the influenced changes of cell proliferation, cell cycle, apoptosis, and soft agar colony formation rate and the migration of transfected cells were analyzed.

RESULTSBefore transfection, the expression level of mature miR-542-3p in 16HBE-T was lower (39.08 ± 6.95)% than it in 16HBE-N (t = 15.18, P < 0.05). In comparison with the 16HBE-T group, the expression level of miR-542-3p in miR-542-3p mimic-transfected group was (5.23 ± 0.55) fold (t = 17.37, P < 0.05) after transfection. Cell proliferation of mimic-transfected group was decreased to (62.06 ± 5.61)% (t = -17.28, P < 0.05), percentage of cells in G(0)/G(1) phase up to (74.76 ± 4.86)% (t = 4.53, P < 0.05), rate of colony formation degrade to (5.87 ± 0.67)% (t = -6.66, P < 0.05), coverage areas ratio decreased to (0.31 ± 0.08) (t = -6.78, P < 0.05). There was no change with apoptosis.

CONCLUSIONOur studies showed that miR-542-3p played the role as a tumor suppressor, which led to a significant decrease in the proliferation capacity and degree of malignancy. These findings suggest aberrantly down-regulated miR-542-3p may be one critical factor that contributes to malignant transformation of 16HBE induced by anti-BPDE.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; adverse effects ; Bronchi ; cytology ; Cell Transformation, Neoplastic ; drug effects ; genetics ; metabolism ; Epithelial Cells ; cytology ; drug effects ; Humans ; MicroRNAs ; genetics ; Transfection

OBJECTIVETo study the inhibitory effect of chlorophyllin (CHL) on trans-benzo(a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide (BPDE) induced malignant transformation in human bronchial epithelial cell line (16HBE).

METHODS10, 50 or 100 micro mol/L CHL were added into the media during the cells transformation induced by BPDE, and the malignant degree of transformed cells were identified by the ConA agglutination test and the assay for anchorage-independent growth and tumorigenicity.

RESULTSAfter the cells were cultured for 25 times, the time of cells agglutination in groups treated with both CHL and BPDE was increased significantly; the colony formation efficiency in soft agar in groups treated with both CHL and BPDE (7.4 per thousand, 11.4 per thousand and 14.4 per thousand ) showed significant decrease (P < 0.05) in dose-dependent manner, as compared with that in group treated with BPDE alone (19.6 per thousand ). Cells treated with both CHL and BPDE or BPDE alone developed tumor in nude mice, a squamous carcinoma confirmed by histopathological examination. The volume of tumor in groups treated with both CHL and BPDE (0.43 +/- 0.13) cm(2), (0.22 +/- 0.04) cm(2) and (0.10 +/- 0.06) cm(3) was significantly smaller (P < 0.05) and dose-dependent, as compared with that in the group treated with BPDE alone (1.71 +/- 0.37) cm(3).

CONCLUSIONCHL showed significant antitransforming ability in human bronchial epithelial cell line induced by BPDE.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; toxicity ; Animals ; Anticarcinogenic Agents ; pharmacology ; Cell Transformation, Neoplastic ; drug effects ; Chlorophyllides ; pharmacology ; Mice ; Mice, Inbred BALB C ; Neoplasms, Experimental ; prevention & control

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; toxicity ; Carcinogens ; toxicity ; Cell Line, Tumor ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; HSP70 Heat-Shock Proteins ; biosynthesis ; Humans

OBJECTIVETo investigate Benzo (a) pyrene (BaP) residue in retail food of Xiamen.

METHODSBaP residue in 121 retail food samples collected from Xiamen were determined by a rapid BaP detector based on derivative constant-energy synchronous fluorescence technique.

RESULTSBaP was detected in 84.3% samples and the concentration were ranged from 0.17 to 59.0 microg/kg. There were 49.6% samples exceeding 5.00 microg/kg, and most of them were roasting food (1.44 - 54.10 microg/kg), processed meat products (0.17 - 59.00 microg/kg) and aquatic products (2.79 - 36.80 microg/kg). The BaP concentration in 34 samples collected from roadside stands were 1.78 - 49.60 microg/kg, of which the rate of the samples exceeding 5.00 microg/kg was 88.2%.

CONCLUSIONThe BaP contamination in retail food samples from Xiamen is serious.

Benzo(a)pyrene ; analysis ; China ; Food Contamination ; analysis ; Food Inspection ; methods ; Meat Products ; analysis