SEARCH · PubMed Health

Results for “GGT”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 163 records · Page 9Linked to original sources

PubMed1997

gamma-Glutamyl transpeptidase-dependent lipid peroxidation in isolated hepatocytes and HepG2 hepatoma cells.

Gamma-glutamyltranspeptidase (GGT), a plasma membrane-bound enzyme, provides the only activity capable to effect the hydrolysis of extracellular glutathione (GSH), thus favoring the cellular utilization of its constituent amino acids. Recent studies have shown however that in the presence of chelated iron prooxidant species can be originated during GGT-mediated metabolism of GSH, and that a process of lipid peroxidation can be started eventually in suitable lipid substrates. The present study was undertaken to verify if a GGT-dependent lipid peroxidation process can be induced in the lipids of biological membranes, including living cells, and if this effect can be sustained by the GGT highly expressed at the surface of HepG2 human hepatoma cells. In rat liver microsomes (chosen as model membrane lipid substrate) exposed to GSH and ADP-chelated iron, the addition of GGT caused a marked stimulation of lipid peroxidation, which was further enhanced by the addition of the GGT co-substrate glycyl-glycine. The same was observed in primary cultures of isolated rat hepatocytes, where the lipid peroxidation process did not induce acute toxic effects. GGT-stimulation of lipid peroxidation was dependent both on the concentration of GSH and of ADP-chelated iron. In GGT-rich HepG2 human hepatoma cells, the exposure to GSH, glycyl-glycine, and ADP-chelated iron resulted in a nontoxic lipid peroxidation process, which could be prevented by means of GGT inhibitors such as acivicin and the serine-boric acid complex. In addition, by co-incubation of HepG2 cells with rat liver microsomes, it was observed that the GGT owned by HepG2 cells can act extracellularly, as a stimulant on the GSH- and iron-dependent lipid peroxidation of microsomes. The data reported indicate that the lipid peroxidation of liver microsomes and of living cells can be stimulated by the GGT-mediated metabolism of GSH. Due to the well established interactions of lipid peroxidation products with cell proliferation, the phenomenon may bear particular significance in the carcinogenic process, where a relationship between the expression of GGT and tumor progression has been envisaged.

Animals↗

PubMed2000

Expression of gamma-glutamyl transpeptidase protects ramos B cells from oxidation-induced cell death.

The ectoenzyme, gamma-glutamyl transpeptidase (GGT, EC ) cleaves glutathione (GSH) to facilitate the recapture of cysteine for synthesis of intracellular GSH. The impact of GGT expression on cell survival during oxidative stress was investigated using the human B cell lymphoblastoid cell line, Ramos. Ramos cells did not express surface GGT and exhibited no GGT enzyme activity. In contrast, Ramos cells stably transfected with the human GGT cDNA expressed high levels of surface GGT and enzymatic activity. GGT-transfected Ramos cells were protected from apoptosis when cultured in cyst(e)ine-deficient medium. The GGT-expressing cells also had lower levels of intracellular reactive oxygen species (ROS). Homocysteic acid and alanine, inhibitors of cystine and cysteine uptake, respectively, caused increased ROS content and diminished viability of GGT expressing cells. Exogenous GSH increased the viability of the GGT-transfected cells more effectively than that of control cells, whereas the products of GSH metabolism prevented death of both the control and GGT-transfected cells comparably. These data indicate that GGT cleavage of GSH and the subsequent recapture of cysteine and cystine allow cells to maintain low levels of cellular ROS and thereby avoid apoptosis induced by oxidative stress.

B-Lymphocytes↗

PubMed2000

Lifestyle and the development of increased serum gamma-glutamyltransferase in middle-aged Japanese men.

To identify the lifestyle factors responsible for increases in serum gamma-glutamyltransferase (GGT), 1014 hepatobiliary dysfunction-free (no medication for and no past history of liver disease, < or = 39 U/L of serum aspartate and/or alanine aminotransferase, and < or = 59 U/L of serum GGT) Japanese male office workers aged 35-55 years were examined annually over five successive years (average period 4.5 years, SD 1.11 years). From the Cox proportional hazards model without serum GGT at entry, significant correlates with the incidence of increased (> or = 60 U/L) serum GGT levels were the slope of body mass index (BMI), alcohol intake, cigarette smoking and coffee drinking (negative). In the model including serum GGT at entry, the slope of BMI and coffee drinking (negative) remained as significant factors for the incidence of increased serum GGT levels. From stepwise regression analyses for the slope of log serum GGT at entry, not including serum GGT in the model, significant correlates with the slope of serum GGT were, in order of relative importance, the slope of BMI, alcohol intake and coffee drinking (negative). In the model with serum GGT at entry, the slope of BMI and coffee drinking (negative) remained as statistically significant. Our results indicate that an increase in body weight is the strongest determinant for increases in serum GGT and that coffee drinking may be associated with a reduced risk of the development of increased serum GGT levels.

Adult↗

PubMed2004

Is serum gamma glutamyltransferase a marker of oxidative stress?

The primary role of cellular gamma glutamyltransferase (GGT) is to metabolize extracellular reduced glutathione (GSH), allowing for precursor amino acids to be assimilated and reutilized for intracellular GSH synthesis. Paradoxically, recent experimental studies indicate that cellular GGT may also be involved in the generation of reactive oxygen species in the presence of iron or other transition metals. Although the relationship between cellular GGT and serum GGT is not known and serum GGT activity has been commonly used as a marker for excessive alcohol consumption or liver diseases, our series of epidemiological studies consistently suggest that serum GGT within its normal range might be an early and sensitive enzyme related to oxidative stress. For example, serum and dietary antioxidant vitamins had inverse, dose-response relations to serum GGT level within its normal range, whereas dietary heme iron was positively related to serum GGT level. More importantly, serum GGT level within its normal range positively predicted F2-isoprostanes, an oxidative damage product of arachidonic acid, and fibrinogen and C-reactive protein, markers of inflammation, which were measured 5 or 15 years later, in dose-response manners. These findings suggest that strong associations of serum GGT with many cardiovascular risk factors and/or events might be explained by a mechanism related to oxidative stress. Even though studies on serum and/or cellular GGT is at a beginning stage, our epidemiological findings suggest that serum GGT might be useful in studying oxidative stress-related issues in both epidemiological and clinical settings.

Biomarkers↗

PubMed1999

Gamma-glutamyl transpeptidase accelerates tumor growth and increases the resistance of tumors to cisplatin in vivo.

We have shown previously that gamma-glutamyl transpeptidase (GGT) activity is essential for the nephrotoxicity of cisplatin. In this study we asked whether GGT activity was necessary for the antitumor activity of cisplatin. GGT was transfected into PC3 cells, a human prostate tumor cell line. Two independent GGT-positive cell lines were isolated and characterized. GGT cleaves extracellular glutathione providing the cells with access to additional cysteine. Expression of GGT had no effect on the growth rate of the cells in vitro where the culture medium contains high levels of cysteine. However, when the cells were injected into nude mice the GGT-positive tumors grew at more than twice the rate of the GGT-negative tumors. Weekly treatment with cisplatin was toxic to both GGT-positive and -negative tumors. The GGT-positive tumors were significantly more resistant to the toxicity of cisplatin than the GGT-negative tumors. Therefore, expression of GGT is required for the nephrotoxicity of cisplatin, but diminishes the tumor toxicity of the drug. These results indicate that the nephrotoxicity and the tumor toxicity of cisplatin are via two distinct pathways.

Animals↗

PubMed1994

Expression of multiple gamma-glutamyl transpeptidase messenger ribonucleic acid transcripts in the adult rat epididymis is differentially regulated by androgens and testicular factors in a region-specific manner.

Multiple gamma-glutamyl transpeptidase (GGT) messenger RNAs (mRNAsII-IV) are expressed in a region-specific manner in the rat epididymis. In the present study, we examined the role(s) of plasma testosterone (T) and testicular factors in regulating the region-specific pattern and quantity of GGT mRNAs expressed along the epididymal duct. Northern blot and ribonuclease protection analyses showed that bilateral orchiectomy for 1, 5, and 15 days dramatically reduced the expression of GGT mRNAsII-IV in the initial segment. Expression of GGT mRNAII and mRNAIII was maintained in the initial segment of orchiectomized animals receiving T implants that maintain normal serum T concentrations, but GGT mRNAIV expression remained low relative to sham-operated control values. Unilateral efferent duct ligation decreased GGT mRNAIV expression only in the initial segment. Hence, expression of GGT mRNAIV in the initial segment was not maintained by circulating T and required a factor(s) of testicular origin. In caput epididymidis, expression of GGT mRNAII and mRNAIII declined after orchiectomy and was not completely restored to control values in orchiectomized animals by plasma T alone, but also required a testicular factor(s). In contrast to the initial segment, expression of GGT mRNAIV in the corpus and cauda epididymidis did not require T and/or a testicular factor(s), as expression of this transcript remained unchanged in these regions after 1, 5, and 15 days of orchiectomy, orchiectomy and T replacement, and after unilateral efferent duct ligation. In the cauda epididymidis, expression of GGT mRNAII required circulating androgens and was unaffected by unilateral efferent duct ligation, whereas GGT mRNAIII expression was repressed by T. These data demonstrate that circulating T and a factor(s) of testicular origin differentially regulate the expression of each GGT mRNA in a region-specific manner.

Androgens↗

PubMed1997

Alteration in gamma-glutamyl transpeptidase activity and messenger RNA of human prostate carcinoma cells by androgen.

Concentrations of the synthetic androgen R1881 that correspond to physiologically relevant concentrations of 5alpha-dihydrotestosterone are capable of altering the activity of gamma-glutamyl transpeptidase (GGT) in human prostate carcinoma cells. GGT activity of the androgen-responsive prostate cancer cell line LNCaP increases >50% above that of the control after a 72-h exposure to 1 nM R1881. This elevation in GGT activity occurs as early as 48 h after treatment and is maintained for at least 96 h. Loss of glutathione (GSH) from media and accumulation of intracellular GSH of cells pretreated with 1 nM R1881 occur at a higher rate than in control cells, suggesting that a greater rate of GSH salvage is associated with the increased GGT activity. Immunohistochemical staining detects an increase in GGT-positive staining in cells treated with 1 nM R1881 for 72 h. Steady-state mRNA levels for GGT are elevated above those of the control 24-72 h after treatment. R1881 has no effect on the GGT activity of the androgen-independent prostate cell line DU145. Growth of LNCaP but not DU145 cells is inhibited by 1 nM R1881 compared to that of the control. Inhibitors of GGT activity, acivicin and serine-borate, are capable of dampening or blocking the effect of R1881 on growth. Growth of LNCaP cells treated with 1 nM R1881 plus 100 mM glycylglycine, a stimulator of GGT activity, is inhibited to a greater extent than the growth of LNCaP cells treated with R1881 alone. These data demonstrate that androgens can elevate GGT activity and increase GGT mRNA and protein levels in human prostate carcinoma cells. In addition, compounds able to alter GGT activity are capable of altering androgen-related growth effects.

Androgens↗

PubMedDate not supplied

Stability and transcriptional regulation of gamma-glutamyl transpeptidase mRNA expression in the initial segment of the rat epididymis.

Gamma-glutamyl transpeptidase (GGT) is an enzyme believed to play a role in the protection of maturing spermatozoa in the epididymis. Our previous studies have shown that four GGT mRNAs (I-IV) transcribed from the single-copy rat GGT gene are differentially expressed and regulated in the rat epididymis. In particular, the normal expression of GGT mRNA(IV) in the epididymal initial segment is dependent upon the presence of testicular factors. The objective of this study was to test the hypothesis that the decreased expression of GGT mRNA(IV) in the initial segment following the in vivo removal of testicular factors by efferent duct ligation (EDL) is due to a decrease in stability and/or transcription rate. The stability of the GGT mRNAs was evaluated by measuring the rate of mRNA decay. These stability studies showed that GGT mRNA(IV) exhibited a rapid initial decay that slowed at later times to a decay rate similar to that of GGT mRNAs(II,III). The decay rates were not different following sham-operation or EDL, and thus the stability of GGT mRNAs were not influenced by the in vivo loss of testicular factors. Results of transcription analysis revealed that the transcription rate of GGT mRNA(IV) in the initial segment fell by approximately 68% following a 12-hour EDL. Additionally, secondary-structure models indicate two families of folding patterns for GGT mRNA(IV), which could be the reason for the two decay regimes detected in the stability study. Thus, the decreased expression level of GGT mRNA(IV) in the initial segment following the in vivo loss of testicular factors is a function of a decreased transcription rate and intricate decay kinetics.

Animals↗

PubMed1992

Repetitive 5-azacytidine treatments of Fao cells induce a stable and strong expression of gamma-glutamyl transpeptidase.

The role of DNA methylation in the expression of the rat gamma-glutamyl transpeptidase (GGT) gene was assessed in the Fao cell line using a hypomethylating agent, 5-azacytidine. Ten repetitive treatments of the cells, with 8 microM 5-azacytidine for 24 h, led to 13- and 80-fold increases, respectively, in GGT activity and in GGT mRNA level. The DNA methylation patterns generated by the isoschizomeric restriction enzymes Hpa II and Msp I indicated that the GGT gene, highly methylated in Fao cells, became strongly demethylated after 5-azacytidine treatments. Thus, DNA demethylation increases the expression of the GGT gene. 5-Azacytidine treatments also increased, but to a lesser extent, mRNAs level for actin, albumin, mitochondrial aspartate aminotransferase, aldolase B mRNAs (12- to 16-fold) as well as for tubulin, gluthathione transferase, and tyrosine aminotransferase mRNAs (2- to 5-fold). The GGT gene expression was further studied in B4 cells, cloned from the demethylated Fao cell population. This clone B4 exhibited a stable and strong GGT activity and a highly demethylated GGT gene. Among the three GGT mRNA I, II, or III, transcribed from three different promoters of the single rat GGT gene, only mRNA III was detected in Fao cells and was increased in clone B4, indicating that the demethylation acts on the promoter for mRNA III. The analysis of the differentiation state of B4 cells, as compared to Fao cells, showed a loss of the regulation of GGT and aspartate aminotransferase genes by dexamethasone, as well as a loss of the gluconeogenic pathway. Interestingly, B4 cells have retained many other specific functions of hepatic differentiation and have acquired alpha-fetoprotein expression; thus this clone exhibits the characteristics of a hepatic fetal phenotype.

Animals↗

PubMed1986

Serum gamma-glutamyl transpeptidase activity in patients receiving chronic phenytoin therapy.

To better define the incidence and range of elevated gamma-glutamyl transpeptidase (GGT) activity in patients taking a drug known to induce hepatic microsomal enzymes, GGT was measured in the sera of 58 patients before and after six months of phenytoin therapy. Enzyme activity at six months was greater than baseline in 52 of 58 patients (90%). GGT activity (normal, 0-50 units/liter) was 45.2 +/- 9.9 (mean +/- SEM) at baseline and 135.8 +/- 18.1 after six months of therapy, a mean threefold increase (P less than 0.001). Of the 45 patients with normal baseline GGT activity, some had marked elevation of serum GGT activity with values rising over 200 units/liter in eight patients and over 300 units/liter in four patients. Mean serum GGT activity remained significantly elevated at 12 and 24 months. This elevation in GGT activity was not influenced by age, sex, or additional anticonvulsant drug therapy. Both baseline and six-month GGT activity was lowest in patients who drank no alcohol, higher in patients who drank 0-1 pint/week, and greatest in patients who drank greater than 1 pint/week. All 13 patients with elevated baseline GGT activity were regular users of alcohol and/or taking other enzyme-inducing drugs. In conclusion, increase in serum GGT activity occurs in 90% of patients on long-term phenytoin therapy, most often to moderate but occasionally to high levels, and this rise in GGT activity is accentuated by regular consumption of alcohol.

Adult↗

PubMed2003

Different mechanisms for gamma-glutamyltransferase-dependent resistance to carboplatin and cisplatin.

In this work, we investigated the effect of gamma-glutamyltransferase (GGT) overexpression on cell viability after carboplatin treatment and compared with cisplatin. Carboplatin challenge of HeLa cells induced GGT and glutamate-cystine ligase (GCL) activities by 2- and 1.4-fold, respectively and concomitantly increased the intracellular reduced glutathione (GSH) level (1.5-fold). To study the role of GGT, HeLa-GGT cells, a stably transfected cell line overexpressing GGT (120-150 mU/mg protein) and the parental HeLa cells (10-15 mU/mg protein) were used. Both cell lines exhibited comparable viability (IC(50) approximately 150 microM) after carboplatin treatment when cultured in standard (250 microM cystine) medium. Culture in low (50 microM) cystine medium resulted in a dramatic decrease (approximately 90%) of the intracellular GSH level and to a 2.5-fold increase of carboplatin cytotoxicity (IC(50) approximately 60 microM). When GSH (50 microM) was included in the culture medium, only HeLa-GGT cells exhibited increased resistance to carboplatin. Using partially purified GGT from HeLa-GGT cells, we show that cisplatin forms adducts with cysteinylglycine, depending only on GGT activity whereas carboplatin did not efficiently react with cysteinylglycine. Thus, in this model system, GGT activity can affect platinum drugs cytotoxocity by two different ways: cisplatin can be detoxified extracellularly after reaction with the -SH group of cysteinylglycine; in the case of carboplatin, the supply of GSH precursors, initiated by GGT, increases the intracellular level of the tripeptide and provides enhanced defensive mechanisms to the cell.

Antineoplastic Agents↗

PubMed2001

Differential regulation of gamma-glutamyltransferase mRNAs in four human tumour cell lines.

Human gamma-glutamyltransferase (GGT) belongs to a multigenic family and at least three mRNAs are transcribed from the gene that codes for an active enzyme. Four human tumour cell lines (HepG2, LNCap, HeLa and U937) with different GGT levels were used to investigate how GGT activity, total GGT mRNA and each individual GGT mRNA subtype responded to tumour necrosis factor-alpha (TNF-alpha), 12-O-tetradecanoylphorbol 13-acetate (TPA) or sodium butyrate treatment. Butyrate reduced the GGT activity in HepG2 cells, and the level of total GGT mRNA accordingly, whereas TNF-alpha and TPA did not alter these parameters. In LNCap cells, TNF-alpha, TPA, and butyrate reduced the activity as well as the level of GGT total mRNA. In HeLa cells no significant changes were observed either in activity or in mRNA level whereas TPA induced both GGT activity and mRNA levels in U937 cells. The distribution of each GGT mRNA subtype (A, B and C) was found to be cell specific: type B mRNA was the major form in HepG2 cells, while type A was the major form in LNCap and HeLa, type A and type C were expressed almost at the same level in U937 cells. The GGT mRNA subtypes were also differently modulated in these cells after TNF-alpha, TPA or butyrate treatment, suggesting that they are regulated by distinct and cell type specific mechanisms.

Butyrates↗

PubMed2002

Fetal gender and cocaine exposure as determinants of cord blood gamma-glutamyl transferase activity.

OBJECTIVE: The serum activity of the hepatic enzyme gamma-glutamyl transferase (GGT) is elevated in the newborn relative to older age groups. Few reports to date have studied the influence of perinatal factors on neonatal serum GGT and no study has assessed the influence of maternal drug ingestion. STUDY DESIGN: Cord blood was randomly collected from 234 liveborn infants and correlated with a range of maternal and fetal perinatal variables to assess influences on cord blood GGT. RESULTS: Our study showed that the range of cord blood GGT activity in 234 randomly selected term newborns was 22 to 556 IU/l. In a subgroup of 75 newborns, GGT activity was independently influenced by only two of the variables studied - cocaine exposure and fetal gender (p=0.009, r=0.39). Females had a lower GGT than males (95+/-66 vs 130+/-90 IU/l, p<0.001) while GGT activity in cocaine-exposed newborns was lower than in cocaine-nonexposed newborns (96+/-48 vs 142+/-109, p<0.01). Birth weight, race, gestational age, and maternal serum GGT were not found to significantly influence cord blood GGT activity. Maternal GGT was uniformly normal and was not affected by any of the variables tested. CONCLUSION: Our findings demonstrate that the reference range for cord blood GGT activity is wide and appears to be influenced by fetal gender and cocaine exposure.

Cocaine↗

PubMed1991

Identification of a human gamma-glutamyl cleaving enzyme related to, but distinct from, gamma-glutamyl transpeptidase.

We have cloned a 2.4-kilobase cDNA from a human placental cDNA library by using a gamma-glutamyl transpeptidase [GGT; gamma-glutamyltransferase, (5-glutamyl)-peptide:amino acid 5-glutamyltransferase, EC 2.3.2.2] probe. The deduced amino acid sequence of this cDNA, GGT-rel, exhibited an overall similarity of 39.5% with human GGT. Sequences that could represent a heavy and a light chain, analogous to GGT, as well as a putative transmembrane region were identified in GGT-rel. Transfectants overexpressing GGT-rel were tested for their ability to catalyze cleavage of the gamma-glutamyl moiety from natural and synthetic substrates for GGT. Experiments with glutathione added to the medium suggested that GGT-rel could hydrolyze the gamma-glutamyl moiety. More definitive evidence was obtained in experiments in which this protein converted leukotriene C4 to leukotriene D4. However, GGT-rel did not convert synthetic substrates that are commonly used to assay GGT. Our results indicate that GGT can no longer be considered the only enzyme capable of cleaving the gamma-glutamyl linkage of leukotriene C4 and, most likely, of other natural compounds.

Amino Acid Sequence↗

PubMed2002

Identification of amino acids important for the catalytic activity of the collagen glucosyltransferase associated with the multifunctional lysyl hydroxylase 3 (LH3).

Collagen glucosyltransferase (GGT) activity has recently been shown to be associated with human lysyl hydroxylase (LH) isoform 3 (LH3) (Heikkinen, J., Risteli, M., Wang, C., Latvala, J., Rossi, M., Valtavaara, M., Myllylä, R. (2000) J. Biol. Chem. 275, 36158-36163). The LH and GGT activities of the multifunctional LH3 protein modify lysyl residues in collagens posttranslationally to form hydroxylysyl and glucosylgalactosyl hydroxylysyl residues respectively. We now report that in the nematode, Caenorhabditis elegans, where only one ortholog is found for lysyl hydroxylase, the LH and GGT activities are also associated with the same gene product. The aim of the present studies is the identification of amino acids important for the catalytic activity of GGT. Our data indicate that the GGT active site is separate from the carboxyl-terminal LH active site of human LH3, the amino acids important for the GGT activity being located at the amino-terminal part of the molecule. Site-directed mutagenesis of a conserved cysteine at position 144 to isoleucine and a leucine at position 208 to isoleucine caused a marked reduction in GGT activity. These amino acids were conserved in C. elegans LH and mammalian LH3, but not in LH1 or LH2, which lack GGT activity. The data also reveal a DXD-like motif in LH3 characteristic of many glycosyltransferases and the mutagenesis of aspartates of this motif eliminated the GGT activity. Reduction in GGT activity was not accompanied by a change in the LH activity of the molecule. Thus GGT activity can be manipulated independently of LH activity in LH3. These data provide the information needed to design knock-out studies for investigation of the function of glucosylgalactosyl hydroxylysyl residues of collagens in vivo.

Amino Acid Sequence↗

PubMed2000

Lectin affinity chromatography and electrophoretic properties of human platelet gamma-glutamyl transferase.

The sialoglycoprotein, gamma-glutamyl transferase (GGT, gamma-GT, EC 2.3.2.2) is a membrane enzyme found in many cells including platelets and leukocytes. In platelets GGT converts leukotriene C4 (LTC4) to leukotriene D4 (LTD4) and is involved in glutathione metabolism. In this study, human platelet GGT was solubilized with Triton X-100 and purified by lectin affinity chromatography on Con A Sepharose 4B to determine its electrophoretic properties. The specific activity of purified GGT was 236 mU/mg protein; 73.7% of human platelet GGT activity was found bound to Con A and 50% of the bound activity was released with 0.3 mol/l methyl alpha-D-mannopyranoside. We observed that human platelet GGT has only one isoenzyme band showing a carbohydrate stained band near the origin on polyacrylamide gel electrophoresis (PAGE). The electrophoretic mobility of papain-solubilized GGT was higher than that of Triton X-100-solubilized GGT at PAGE. Also GGT activities were determined on neuraminidase, trypsin or n-butanol-DIPE (diisopropyl ether)-treated Triton X-100-solubilized membrane fractions. This characterization may be useful when trying to establish the contribution of platelet GGT to serum GGT activity. This marker may reflect the extent of platelet activation.

Adult↗

PubMed1990

gamma-Glutamyltranspeptidase-positive rat hepatocytes are protected from GSH depletion, oxidative stress and reversible alterations of collagen receptors.

The aim of this study has been to define cytotoxic mechanisms that may cause clonal expansion in the liver of pre-carcinogenic cells. An in vitro model, which has been described previously, was used. Hepatocytes were isolated from carcinogen-treated rats and a high proportion of the cells were gamma-glutamyltranspeptidase (GGT)-positive. The cells were incubated in suspension and exposed to toxic agents in concentrations that induced a moderate increase in cellular leakage within 3 h. Samples were withdrawn and sampled cells were then allowed to attach to collagen-coated plates. Attached cells were stained and the ratio of GGT-positive/GGT-negative cells (GGT-ratio) was determined. The initial GGT-ratio was 10.4 +/- 4.7% and an increased ratio was taken as a sign of toxicity that resulted in a selection of GGT-positive cells. In a first series of experiments it was shown that hydroquinone and menadione increase the GGT-ratio, while diquat, sodium selenite, diethyl maleate or phorone do not. However, diethyl maleate in combination with diquat increased the GGT-ratio. Hydrogen peroxide (5 mM) increased the GGT-ratio as effectively as hydroquinone (0.3 mM). Lower concentrations of H2O2 (0.05 mM) increased the GGT-ratio in GSH-depleted cells. The changes induced by hydroquinone and H2O2 in low concentration were reversible. In another series of experiments, plates coated with antibodies against beta 1-integrin were used. An increase in the GGT-ratio was obtained with anti beta 1-integrin, but not with broad spectrum anti-rat hepatocyte or anti-rat beta 2-microglobulin antibodies as substrata. These data suggested an involvement of the beta 1-integrin in the selection. Taken together, these data indicate that GGT-positive hepatocytes are protected against GSH depletion and oxidative stress that may result in reversible receptor alterations.

Animals↗

PubMed1993

Induction of three histochemically distinct populations of hepatic foci in C57BL/6J mice.

Although expression of the enzyme gamma-glutamyl transpeptidase (GGT) is the most common phenotypic marker of preneoplastic foci in the livers of carcinogen-treated rats, it is not generally expressed in mouse liver tumors or hepatic foci. However, several carcinogens, including safrole and ortho-azoaminotoluene (OAT), have been reported to induce GGT-positive foci in mice. We asked whether safrole and OAT induce GGT expression in preneoplastic foci or if these compounds select for a distinct set of lesions that can be identified by their GGT-positive phenotype. We treated 12-day-old male and female C57BL/6J mice with N,N-diethylnitrosamine (DEN) (0.20 mumol/g body wt) to initiate hepatocarcinogenesis. From 6 to 24 weeks of age, during the promotion phase of hepatocarcinogenesis, groups of mice were treated with 3,4,5,3',4',5'-hexabromobiphenyl (HBB), safrole or OAT. Additional groups of female mice were ovariectomized at 6 weeks of age with or without subsequent chronic treatment with testosterone. All the animals were killed at 24 weeks of age and serial liver sections were stained for glucose-6-phosphatase (G6Pase) or GGT. Both testosterone and HBB were strong promoters of the development of G6Pase-deficient foci. No GGT-positive foci were observed in animals treated with these agents or with DEN alone. In mice fed safrole or OAT during the promotion period, female mice developed more G6Pase-deficient foci than male mice, and GGT-positive foci were observed. Analysis of serial sections revealed that the G6Pase-deficient foci and the GGT-positive foci were independent populations. The relative number of these two classes of foci varied according to the treatment regimen. In females fed safrole, 7% of the foci in the liver were GGT-positive while in female mice fed OAT, 45% were GGT-positive. In all groups of mice in which we observed GGT-positive foci and in ovariectomized female mice, we noted a third independent population of foci which demonstrated significantly increased expression of G6Pase relative to surrounding normal liver. These data indicate that different treatments during the promotion stage of hepatocarcinogenesis in the mouse may give rise to distinct populations of preneoplastic lesions. Further studies of the molecular events giving rise to these distinct lesions will provide insights into the multiple pathways that result in hepatocarcinogenesis.

Animals↗