OBJECTIVETo investigate the modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1-hydroxypyrene (1-OHP) excretions in workers under different exposure levels.

METHODSFour hundred and forty-seven occupationally exposed workers from two coking plants and 220 control workers from a wire rod plant were genotyped to analyze the modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1-OHP excretions.

RESULTSThe urinary 1-OHP concentration in exposed group was much higher than that in control group (4.61 vs 0.34 µmol/mol Cr, P < 0.05). Occupational exposure levels and cigarette smoking were of the dominating factors affecting 1-OHP excretions in urine. After controlling potential confounders, decreased excretion of urinary 1-OHP was associated with GSTP1 I105V AG + GG genotype in coke oven workers (single-gene model, P = 0.012; multi-gene model, P = 0.011) and with GSTT1 null type in the analysis including all subjects (P = 0.055 in both single-gene and multi-gene models). GSTT1 and GSTP1 were interacted on the urinary concentrations of 1-OHP.

CONCLUSIONUrinary 1-OHP concentrations can be modified by GSTM1, GSTT1 and GSTP1 gene polymorphisms, indicating that these genes are involved in the metabolism of polycyclic aromatic hydrocarbons.

Adolescent ; Adult ; Control Groups ; Genotype ; Glutathione S-Transferase pi ; genetics ; Glutathione Transferase ; genetics ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Polymorphism, Single Nucleotide ; Pyrenes ; analysis ; Urinalysis ; Young Adult

To identify the inhibitor of glutathione S-transferase (GST), a high-throughput screening method was established in a 384-well microplate with total 35 microL volume, and the absorbance at 340 nm is detected. The concentrations of substrates, CDNB and GST were determined by chromatometry. The optimal enzyme kinetics reaction time and temperature are 2 h and 30 degrees C , respectively. The established model was evaluated by NaOCl, a known GST inhibitor, and the parameter Z' was 0.77, which showed a high feasibility and stability of the assay. A total of 31,098 compounds were screened, of which 4 compounds were shown to inhibit GST activity, high inhibiting activity for their IC50 of GST inhibition was 3.94, 4.05, 74.85, and 77.41 mg x L(-1), separately. The results indicated that the colorimetric method by using CDNB and GSH as substrate is stable, sensitive, reproducible and also suitable for high throughput screening.
Dinitrochlorobenzene ; chemistry ; Drug Evaluation, Preclinical ; methods ; Enzyme Inhibitors ; analysis ; Glutathione ; chemistry ; Glutathione Transferase ; antagonists & inhibitors ; Substrate Specificity

OBJECTIVETo study the relationship between glutathione S-transferase M1 (GSTM1) genotypes and lipid peroxidation of asbestos workers.

METHODS94 asbestos workers and 51 controls were selected as subjects. The general information, occupational history and individual habits were collected by questionnaires in all participants. The venous blood was sampled and the plasma was separated for the detection of malondialdehyde (MDA) level and lymphocytes for DNA isolation and GSTM1 genotyping.

RESULTSMDA level was significantly higher in asbestos workers [(0.283 +/- 0.054) nmol/L] than that in controls [(0.163 +/- 0.053) nmol/L, P < 0.01], however, neither duration of exposure nor accumulated asbestos exposure dose was related to MDA levels; MDA levels in control workers with GSTM1 +/- genotype [(0.190 +/- 0.034) nmol/L] were significantly higher than that in control workers with GSTM1 +/+ genotype [(0.138 +/- 0.055) nmol/L, P < 0.01]. Among asbestos workers, the same trend could be found, but the differences was not significant(P > 0.05). When the workers were stratified by duration of exposure or accumulated asbestos exposure dose, MDA levels in individuals with GSTM1 -/- genotype were also higher than those with GSTM1 +/+ genotype, but the differences were also not significant(P > 0.05).

CONCLUSIONBoth exposure to asbestos and deficiency of GSTM1 genotype were related to lipid peroxidation in workers, but the role of the former may be more important than that of the latter.

Asbestos ; adverse effects ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Lipid Peroxidation ; Malondialdehyde ; analysis ; Occupational Exposure

OBJECTIVESTo identify hepatic progenitor cells (HPCs) in patients with severe hepatitis (SH) by detecting their markers and investigate the features of their distribution and location.

METHODSLiver tissues taken from 59 SH patients were tested for the receptor of stem cell factor (c-kit), pi-class glutathione S-transferase (GST-pi), cluster of differentiation 34 (CD34), cytokeratin 19 (CK19), cytokeratin 18 (CK18) and alpha fetoprotein (AFP) by immunohistochemistry (IHC). Meanwhile, 58 patients with acute or chronic hepatitis were also detected to act as controls.

RESULTSHepatic progenitor cells could be seen in SH patients. Most of them existed as ductular cells that had been called "typical ductular proliferation (ADP)" or "typical ductular reaction" in previous research. These ductular cells were mainly located at the portal areas, fibro septa, periportal parenchyma and the border of the pseudolobuli and inflammatory foci. Further, c-kit, GST-pi, CK19 and CK18, but not CD34 and AFP could be detected in these cells. Another kind of HPC was the small hepatocyte-like cell (SHLC), which could express c-kit, GST-pi, and CK18, but not CK19, CD34 and AFP. The semi-quantitative analysis showed that the scope of ADP in SH patients was significantly larger than that in acute and chronic hepatitis patients (chi2= 63.62, P<0.05), and the scope of ADP in subacute severe hepatitis and chronic severe hepatitis patients was also significantly larger than that in acute severe hepatitis patients.

CONCLUSIONIn the course of regeneration of viral hepatitis, different types of pathology have different features. In acute and chronic hepatitis (G1-2), the regeneration is mainly owing to the proliferation of mature hepatocytes, and in chronic hepatitis (G3-4), there is the participation of HPCs, although they are limited. In severe hepatitis, however, since the replicative capacity of normal hepatocytes is impaired or prohibited, liver regenerates and restores mainly by the means of hepatic stem cells activation and proliferation. But the hepatic stem cells don't differentiate into their mature functional compartments directly at all. There are several intermediary or transition populations. In human severe hepatitis, they are mainly ductular cells, and parts of them are small hepatocyte-like cells.

Antigens, CD34 ; analysis ; Cell Division ; Female ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Hepatitis ; pathology ; Hepatocytes ; pathology ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Keratins ; analysis ; Male ; Proto-Oncogene Proteins c-kit ; analysis ; Stem Cells ; pathology ; alpha-Fetoproteins ; analysis

BACKGROUND: P-glycoprotein (PGP) is capable of expelling cytotoxic drugs from cytosol and the overexpression mediates drug resistance. However not all resistant leukemic cells express PGP. High expression of glutathione S-transferasepie (GSTpie) is related to clinical outcome following chemotherapy. Topoisomerase IIalpha (topo IIalpha) is a major target of anthracyclines for the treatment of leukemia. METHODS: To evaluate the relation of PGP, GSTpie and topo IIalpha expression to treatment outcome, PGP, GSTpie and topo IIalpha expression were analysed by flow cytometry using mono clonal antibodies (anti-JSB1, anti-GSTpie and anti-topo IIalpha) in 33 cases of de novo acute myelogenous leukemia. RESULTS: In patients with AML, the frequency of patients with high expression of PGP was 57.6% (19/33). The complete remission (CR) rate and mean survival duration were significantly different between patients with high expression and those with low expression of PGP (31.6 vs 92.9%, P=0.001; 83 vs 341 days, P=0.011). The frequency of patients with high expression of GST pie was 60.6% (20/33). The CR rate and mean survival duration were significantly different between patients with high expression and those with low expression of GSTpie (40.0 vs 84.6%, P=0.011; 115 vs 343 days, P=0.021). The frequency of patients with high expression of topo IIalpha is 78.8% (26/33) and treatment outcome was not related to topo IIalpha expression. In multivariate analysis with age, WBC count, PGP and GSTpie, PGP expression was an independent prognostic factor for treatment outcome. CONCLUSION: The flow cytometric measurement of PGP and GSTpie expression can be useful for the prediction of treament outcome following chemotherapy and PGP can be used as aprognostic factor in AML.
Adult* ; Anthracyclines ; Antibodies ; Cytosol ; DNA Topoisomerases, Type II* ; Drug Resistance ; Drug Resistance, Multiple ; Drug Therapy ; Flow Cytometry ; Glutathione S-Transferase pi* ; Glutathione Transferase* ; Glutathione* ; Humans ; Leukemia ; Leukemia, Myeloid, Acute* ; Multivariate Analysis ; P-Glycoprotein* ; Treatment Outcome

An experiment was conducted in order to investigate the effect of p-dimethylaminoazobenzene (DAB) and 2(3)-tert-butyl-4-hydroxyanisole (BHA) on the lipid peroxidation and peroxide-destroying enzyme system in the rat liver. Dietary supplementation of DAB (0.06%) for three weeks caused the elevation of glutathione-S-transferase activity by 60% and glutathione reductase by 50%, but it decreased glutathione peroxidase and catalase activities significantly. Dietary supplementation of BHA (0.75%) also increased glutatione-S-transferase activity in the liver by 2 folds, and it counteracts DAB effect on the glutathione peroxidase and catalase activities. There was a marked increase in malon-dialdehyde content in the postnuclear fraction of liver by the treatment of DAB, but the addition of BHA lowered the malondialdehyde content to almost the control level. The protective effect of BHA on the lipid peroxidation induced by DAB administration at the enzyme level seems to be due to the induction of glutathione-S-transferase and the protection of glutathione peroxidase and catalase activities from being lowered by DAB administration.
Animal ; Anisoles/pharmacology* ; Butylated Hydroxyanisole/pharmacology* ; Glutathione Peroxidase/analysis* ; Glutathione Reductase/analysis* ; Glutathione Transferase/analysis* ; Lipid Peroxides/metabolism* ; Liver/drug effects* ; Liver/metabolism ; Male ; Peroxidases/analysis* ; Rats ; p-Dimethylaminoazobenzene/pharmacology*

PURPOSE: Cancer development depends on not only activation of oncogene or inactivation of tumor suppressor gene but also activities of enzymes involved in the metabolism of various carcinogenic xenobiotics, such as arylamine N-acetyltrasferase 2(NAT 2) and glutathione S-transferase (GSTM1). We analyzed whether genetic polymorphisms of NAT2 and GSTM1 were correlated with the mutation patterns of p53 and H-ras genes in bladder tumor tissues. MATERIALS AND METHODS: In 49 bladder cancer patients, we performed direct DNA sequencing for the detection of mutations of p53 and H-ras gene in bladder tumor tissues, and adopted PCR and PCR-RFLP techniques for the analysis of genetic polymorphisms of NAT2 and GSTM1 using patients` blood samples, respectively. RESULTS: In 18 cases, mutations in p53 were detected whereas 1 case carried two mutations; thus total of 19 mutations were detected. Sixteen of these were point mutations including 13 of transversions and 3 of transitions, and others were 1 of frameshift and 2 of microdeletions-insertions. Among 33 patients, H-ras mutations were detected in 5 cases with 2 of transitions and 3 of transversions. The frequencies of slow, intermediate, and rapid acetylator in NAT2 genotyping analysis, were 10.2%, 40.8%, and 49.0%, respectively, and GSTM1 deletions were observed in 73.5%. We could not find any significant correlations between NAT2 or GSTM1 polymorphisms and the occurence of p53(p=0.614, p=0.310) or H-ras(p=0.500, p=0.582) mutations. Also, no apparent associations were seen for specific type of p53 and H-ras mutations according to polymorphisms of NAT2(p=0.456, p=0.600) and GSTM1(p=0.378, p=0.400). CONCLUSIONS: The polymorphisms of NAT2 and GSTM1, conjugating enzymes of foreign compound metabolism, were not considered to influence occurrence and type of mutations in p53 and H-ras in human bladder cancer.
Genes, ras ; Genes, Tumor Suppressor ; Glutathione Transferase ; Humans* ; Metabolism ; Oncogenes ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Urinary Bladder Neoplasms* ; Urinary Bladder* ; Xenobiotics

OBJECTIVETo study the relationship between glutathione S transferase M1(GST mu) gene deletion and leukemia in workers exposed to benzene.

METHODSA matched population-based case-control survey with multivariate Logistic regression analysis was conducted in this study.

RESULTSIn the population of 34 patients and their matched controls, the absence of the GST mu genotype conferred odds ratio of 3.6. It suggested that GST mu was an important determinant of heterogeneity in individual susceptibility to leukemia associated with exposure to benzene. The single-variance analysis indicated that these markedly significant factors were GST mu gene deletion, GST mu isoenzyme activity, duration of exposure, GST isoenzyme activity, smoking quantity and average concentration of benzene in workshop air. The multivariate analysis indicated that these markedly significant factors were GST mu gene deletion, duration of exposure to benzene and GST mu isoenzyme activity.

CONCLUSIONGST mu gene deletion may be associated with increased risk of leukemia in workers exposed to benzene and is one of genetically determined factors.

Benzene ; toxicity ; Case-Control Studies ; Gene Deletion ; Glutathione Transferase ; genetics ; Humans ; Leukemia ; enzymology ; etiology ; genetics ; Logistic Models ; Multivariate Analysis ; Occupational Exposure ; adverse effects

The aim of this study is to develop monoclonal antibody against human hepatocyte growth factor activator inhibitor 1 (HAI-1) for future study of HAI-1. The cDNA fragments of human hepatocyte growth factor activator inhibitor 1 (HAI-1) were subcloned to construct GST-HAI-1 fusion protein expression vectors. The vectors were transformed into E. coli and fusion protein expression was induced by IPTG. The GST-HAI-1 fusion proteins were separated on preparative SDS-PAGE and recovered by electroelution, and used to immunize BALB/c mice. Hybridomas producing monoclonal antibodies against human HAI-1 were prepared by cell fusion technique and characterized by ELISA, Western Blot and immunohistochemical staining. One hybridoma cell line, ZMC6, was obtained, which produces specific antibody against the expressed GST-HAI-1 fusion protein. The monoclonal antibody recognizes both the membrane-type and secretory-type HAI-1 proteins of colorectal tissue. The successful development of anti-HAI-1 antibody provides a powerful tool for further investigation on HAI-1's function.
Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Glutathione Transferase ; genetics ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Proteinase Inhibitory Proteins, Secretory ; analysis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; immunology

OBJECTIVEThe aim of this study was to investigate the mechanism(MDR) of multidrug resistance(MDR) of mucoepidermoid carcinoma in salivary gland.

METHODS40 cases of mucoepidermoid carcinoma in salivary gland were examined the MDR gene product P-glycoprotein using a monoclonal antibody JSB-1. And 10 of them were also investigated by detecting the expression of GST-pi. All the cases had not been accepted any therapy before the samples were collected.

RESULTS1. Positive expression of JSB-1 was observed in 27 of the 40 specimens. The positive expression was related not only with clinical stage, but also with differentiation degree. 2. The GST-pi positive expression was found in 9 of 10 cases. There was no significant different between the positive expression of JSB-1 and GST-pi.

CONCLUSIONJSB-1 and GST-pi play an important role in MDR of mucoepidermoid carcinoma.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Adolescent ; Adult ; Aged ; Antibodies, Monoclonal ; analysis ; Carcinoma, Mucoepidermoid ; immunology ; Child ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Genes, MDR ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Humans ; Isoenzymes ; analysis ; Male ; Middle Aged ; Salivary Gland Neoplasms ; immunology ; Salivary Glands ; immunology