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1.
Aquat Toxicol ; 170: 1-12, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26562184

ABSTRACT

Nickel compounds are widely used in industries and have been massively introduced in the environment in different chemical forms. Here we report the effect of two different chemical forms of nickel, NiCl2 and nickel nanoparticles (NiNPs), on the reproduction of the marine calanoid copepod Acartia tonsa. The behavior of nickel nanoparticles was analyzed with different techniques and with two protocols. In the "sonicated experiment" (SON) NiNP solution was sonicated while in the "non-sonicated experiment" (NON-SON) the solution was vigorously shaken by hand. Final nominal concentrations of 5, 10 and 50mgL(-1) and 1, 5 and 10mgL(-1) NiNPs were used for the acute and semichronic tests, respectively. Nanoparticle size did not change over time except for the highest concentration of 50mgL(-1) NiNPs, in which the diameter increased up to 843nm after 48h. The concentration of Ni dissolved in the water increased with NP concentration and was similar for SON and NON-SON solutions. Our results indicate that sonication does not modify toxicity for the copepod A. tonsa. Mean EC50 values were similar for NON-SON (20.2mgL(-1)) and SON experiments (22.14mgL(-1)) in the acute test. Similarly, no differences occurred between the two different protocols in the semichronic test, with an EC50 of 7.45mgL(-1) and 6.97mgL(-1) for NON-SON and SON experiments, respectively. Acute and semichronic tests, conducted exposing A. tonsa embryos to NiCl2 concentrations from 0.025 to 0.63mgL(-1), showed EC50 of 0.164 and 0.039mgL(-1), respectively. Overall, A. tonsa is more sensitive to NiCl2 than NiNPs with EC50 being one order of magnitude higher for NiNPs. Finally, we exposed adult copepods for 4 days to NiCl2 and NiNPs (chronic exposure) to study the effect on fecundity in terms of daily egg production and naupliar viability. Egg production is not affected by either form of nickel, whereas egg viability is significantly reduced by 0.025mgL(-1) NiCl2 and by 8.5mgL(-1) NiNPs. At NiNP concentration below the acute EC50 (17mgL(-1)) only 9% of embryos hatched after 4 days. Interestingly, the percentage of naupliar mortality (>82%) observed in the semichronic test at the nominal concentration of 10mgL(-1) NiNPs corresponding to almost 0.10mgL(-1) of dissolved Ni, was similar to that recorded at the same Ni salt concentration. Electron microscopical analyses revealed that A. tonsa adults ingest NiNPs and excrete them through fecal pellets. To the best of our knowledge, this is the first study investigating the toxicity of two different forms of Ni on the reproductive physiology of the copepod A. tonsa and showing the ability of the calanoid copepod to ingest nanoparticles from seawater.


Subject(s)
Aquatic Organisms/drug effects , Copepoda/drug effects , Nanoparticles/toxicity , Nickel/toxicity , Water Pollutants, Chemical/toxicity , Animals , Biological Assay , Female , Nanoparticles/ultrastructure , Particle Size , Spectrophotometry, Atomic , Static Electricity , Toxicity Tests, Acute , Toxicity Tests, Chronic
2.
Cell Death Dis ; 6: e1705, 2015 Mar 26.
Article in English | MEDLINE | ID: mdl-25811802

ABSTRACT

Many developing tissues display regenerative capability that allows them to compensate cell loss and preserve tissue homeostasis. Because of their remarkable regenerative capability, Drosophila wing discs are extensively used for the study of regenerative phenomena. We thus used the developing wing to investigate the role played in tissue homeostasis by the evolutionarily conserved eukaryotic H/ACA small nucleolar ribonucleoprotein pseudouridine synthase. Here we show that localized depletion of this enzyme can act as an endogenous stimulus capable of triggering apoptosis-induced proliferation, and that context-dependent effects are elicited in different sub-populations of the silenced cells. In fact, some cells undergo apoptosis, whereas those surrounding the apoptotic foci, although identically depleted, overproliferate. This overproliferation correlates with ectopic induction of the Wg and JAK-STAT (Janus kinase-signal transducer and activator of transcription) mitogenic pathways. Expression of a p35 transgene, which blocks the complete execution of the death program and generates the so-called 'undead cells', amplifies the proliferative response. Pseudouridine synthase depletion also causes loss of apicobasal polarity, disruption of adherens cell junctions and ectopic induction of JNK (c-Jun N-terminal kinase) and Mmp1 (matrix metalloproteinase-1) activity, leading to a significant epithelial reorganization. Unexpectedly, cell-nonautonomous effects, such as epithelial mesenchymal transition in the contiguous unsilenced squamous epithelium, are also promoted. Collectively, these data point out that cell-cell communication and long-range signaling can take a relevant role in the response to pseudouridine synthase decline. Considering that all the affected pathways are highly conserved throughout evolution, it is plausible that the response to pseudouridine synthase depletion has been widely preserved. On this account, our results can add new light on the still unexplained tumor predisposition that characterizes X-linked dyskeratosis, the human disease caused by reduced pseudouridine synthase activity.


Subject(s)
Apoptosis/genetics , Drosophila Proteins/biosynthesis , Drosophila Proteins/genetics , Epithelial-Mesenchymal Transition/genetics , Hydro-Lyases/genetics , Intramolecular Transferases/genetics , Nuclear Proteins/genetics , Wnt1 Protein/biosynthesis , Animals , Cell Proliferation/genetics , Drosophila Proteins/antagonists & inhibitors , Drosophila melanogaster/genetics , Drosophila melanogaster/growth & development , Gene Expression Regulation, Developmental , Humans , Hydro-Lyases/antagonists & inhibitors , Intramolecular Transferases/antagonists & inhibitors , JNK Mitogen-Activated Protein Kinases/biosynthesis , JNK Mitogen-Activated Protein Kinases/genetics , Matrix Metalloproteinase 1/biosynthesis , Matrix Metalloproteinase 1/genetics , Nuclear Proteins/antagonists & inhibitors , RNA-Binding Proteins , Regeneration/genetics , Signal Transduction , Wings, Animal/enzymology , Wings, Animal/growth & development , Wnt1 Protein/genetics
3.
J Endocrinol Invest ; 35(4): 407-12, 2012 Apr.
Article in English | MEDLINE | ID: mdl-21737999

ABSTRACT

BACKGROUND: Obesity and insulin resistance predispose individuals to the development of both metabolic syndrome and non-toxic nodular thyroid diseases. AIM: The aim of this observational, cross-sectional study is to evaluate the relationship between metabolic syndrome and multinodular nontoxic goiter in an inpatient population from a geographic area with moderate iodine deficiency. SUBJECTS AND METHODS: We examined 1422 Caucasian euthyroid inpatients. Thyroid volume was determined by ultrasound of the neck. A fine-needle aspiration biopsy was performed to evaluate single thyroid nodules and dominant nodules ≥15 mm in euthyroid multinodular goiter. The diagnosis of metabolic syndrome was made according to the criteria of the American Heart Association/ National Heart, Lung, and Blood Institute. RESULTS: Of the sample, 277 patients had clinical evidence of multinodular nontoxic goiter, 461 met the criteria for the diagnosis of metabolic syndrome, and 132 were found to have both conditions. After adjusting for age, gender, body mass index, nicotinism, parity, alcohol intake, thyroid function, and metabolic syndrome- related pharmacological treatment, metabolic syndrome was found to be an independent risk factor for the occurrence of multinodular non-toxic goiter. The relationship between metabolic syndrome and multi nodular non-toxic goiter was apparent in both men and women. CONCLUSIONS: In this study of euthyroid inpatients, we demonstrate that metabolic syndrome is an independent risk factor for the occurrence of multinodular non-toxic goiter in a geographic area with moderate iodine deficiency. We propose that patients meeting the criteria for metabolic syndrome should be screened for the presence of multinodular non-toxic goiter.


Subject(s)
Goiter, Nodular/blood , Goiter, Nodular/epidemiology , Hospitalization , Iodine/deficiency , Metabolic Syndrome/blood , Metabolic Syndrome/epidemiology , Aged , Aged, 80 and over , Female , Humans , Iodine/blood , Italy/epidemiology , Male , Middle Aged , Risk Factors
4.
Bone Marrow Transplant ; 35(4): 323-34, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15543194

ABSTRACT

Cardiac toxicity is an uncommon but potentially serious complication of high-dose (HD) chemotherapy and little is known about incidence, severity and underlying mechanisms. We have systematically reviewed the literature of the last 30 years to summarize and appraise the published evidence on cardiac toxicity associated with HD chemotherapy. HD cyclophosphamide-containing regimens have been most commonly associated with cardiac toxicity, with a progressively decreasing incidence over time. Dosage, application regimens and coadministration of other chemotherapeutic agents emerged as risk factors. While cardiac toxicity has been rarely associated with other cytotoxic drugs, an unexpected incidence of severe cardiotoxicity resulted from reduced-intensity conditioning regimens containing melphalan and fludarabine. Predictive value of cardiologic examination of patients is limited, and patients with a slight depression of cardiac performance could tolerate HD chemotherapy. Clinical examination, resting electrocardiography and dosage adjustment in overweight patients remain the mainstay of prevention, with bidimensional echocardiography (2D echo) for patients with a history of anthracycline exposure. Strategies to decrease the long-term negative impact of anthracycline administration on cardiac performance are being investigated. New 2D echo-based techniques and circulating markers of cardiac function hold promise for allowing identification of patients at high risk for and early diagnosis of cardiac toxicity.


Subject(s)
Antineoplastic Agents/toxicity , Electrocardiography/drug effects , Heart Diseases/chemically induced , Heart Diseases/diagnosis , Humans , Risk Factors
5.
Oncogene ; 20(43): 6132-41, 2001 Sep 27.
Article in English | MEDLINE | ID: mdl-11593421

ABSTRACT

The HMGI proteins (HMGI, HMGY and HMGI-C) have an important role in the chromatin organization and interact with different transcriptional factors. The HMGI genes are expressed at very low levels in normal adult tissues, whereas they are very abundant during embryonic development and in several experimental and human tumours. In order to isolate proteins interacting with the HMGI(Y) proteins, a yeast two-hybrid screening was performed using the HMGI(Y) protein as bait. This analysis led to the isolation of homeodomain-interacting protein kinase-2 (HIPK2), a serine/threonine nuclear kinase. HIPK2 co-immunoprecipitates with the HMGI(Y) protein in 293T cells. The interaction between HIPK2 and HMGI(Y) occurs through the PEST domain of HIPK2 and it is direct because in vitro translated HIPK2 binds HMGI(Y). We also show that HIPK2 is able to phosphorylate the HMGI(Y) protein by an in vitro kinase assay. In order to understand a possible role of HIPK2 gene in cell growth we performed a colony assay which showed an impressive HIPK2 inhibitory effect on normal thyroid cells. Flow cytometric analysis would indicate the block of cell growth at the G2/M phase of the cell cycle. Since normal thyroid cells do not express detectable HMGI(Y) protein levels, we assume that the HIPK2 inhibitory effect is independent from the interaction with the HMGI(Y) protein.


Subject(s)
Carrier Proteins/metabolism , High Mobility Group Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , Transcription Factors/metabolism , Binding Sites , Bromodeoxyuridine/metabolism , Carrier Proteins/chemistry , Carrier Proteins/genetics , Catalysis , Cell Division/drug effects , Cell Line , Cell Nucleus/enzymology , DNA, Complementary/metabolism , Flow Cytometry , Gene Library , HMGA1a Protein , High Mobility Group Proteins/chemistry , High Mobility Group Proteins/genetics , Humans , Models, Genetic , Phosphorylation , Plasmids/metabolism , Precipitin Tests , Protein Binding , Protein Biosynthesis , Protein Serine-Threonine Kinases/chemistry , Protein Serine-Threonine Kinases/genetics , Protein Structure, Tertiary , Recombinant Fusion Proteins/metabolism , Transcription Factors/chemistry , Transcription Factors/genetics , Transfection , Two-Hybrid System Techniques
6.
Bone Marrow Transplant ; 28(3): 277-82, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11535996

ABSTRACT

High-dose cyclophosphamide (HD-CTX) is largely employed in high-dose chemotherapy (HD-CHT) protocols. HD-CTX dose-limiting toxicity expresses itself as cardiac toxicity which is fatal in a minority of patients. The pathophysiology of HD-CTX-associated cardiotoxicity is still poorly understood. Autopsy studies in patients who died from acute HD-CTX-induced cardiac toxicity revealed hemorrhagic myocardial cell death and interstitial edema. Recently troponins, in particular troponin I (cTnI), have been found to represent a uniquely sensitive and specific marker of myocyte membrane integrity and therefore to increase in response to minimal myocardial cell damage in different settings, including doxorubicin-induced cardiotoxicity. We performed a multiparametric cardiologic monitoring in 16 consecutive breast cancer patients undergoing HD-CTX by means of serial ECG registrations and cardiac enzymes (CPK, CPK-MB and cTnI) determinations plus echocardiography in order to clarify acute cardiac events following HD-CTX administration. Neither overt cardiac toxicity nor cardiac enzymes elevation were recorded. Serial ECGs revealed in six cases little and reversible reduction of QRS voltage and/or ST abnormalities. Echo monitoring showed in four cases mild and transient increase of LV diastolic/systolic diameter/volume without decrease of FS% or EF% below normal values: in two of them abnormalities of diastolic function (E/A mitral doppler ratio) were also recorded. We conclude that our protocol of HD-CTX administration does not cause myocardial cell damage as analyzed by serum cTnI levels, thus suggesting that myocyte membrane injury may not be the first direct mechanism of HD-CTX cardiotoxicity. ECG (ie QRS voltages ) and Echo (ie E/A ratio) monitoring leads us to hypothesize that slight interstitial edema with reduction of LV diastolic compliance may be initial signs of cardiac dysfunction in this clinical setting.


Subject(s)
Antineoplastic Agents, Alkylating/toxicity , Breast Neoplasms/drug therapy , Cyclophosphamide/toxicity , Electrocardiography/drug effects , Troponin I/blood , Adult , Antineoplastic Agents, Alkylating/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Biomarkers/blood , Breast Neoplasms/complications , Cyclophosphamide/administration & dosage , Dose-Response Relationship, Drug , Female , Heart Diseases/blood , Heart Diseases/chemically induced , Heart Diseases/diagnosis , Hematopoietic Stem Cell Transplantation/adverse effects , Humans , Middle Aged , Transplantation, Autologous/adverse effects
7.
Ital Heart J Suppl ; 1(11): 1457-63, 2000 Nov.
Article in Italian | MEDLINE | ID: mdl-11109196

ABSTRACT

BACKGROUND: High-dose cyclophosphamide is the nucleus for virtually all high-dose chemotherapy protocols. Non-hematologic dose-limiting toxicity is represented by acute cardiomyopathy, even fatal in a minority of patients. The pathophysiology of such a cardiotoxicity is still poorly understood. Postmortem studies revealed hemorrhagic myocardial cell death, endothelial damage, and interstitial edema. Recently troponins, in particular troponin I, have been found to represent uniquely sensitive and specific markers of myocyte membrane integrity, thus to increase in response to myocardial cell damage in different clinical settings. METHODS: We performed a multiparametric monitoring in 16 consecutive breast cancer patients undergoing cyclophosphamide, by means of serial ECGs, cardiac enzymes determinations (creatine phosphokinase, MB mass and troponin I) through 0 to 72 hours, and echocardiography at baseline and after 48 hours. RESULTS: Neither overt cardiac failure nor enzyme elevation were recorded. Serial ECGs revealed a reduction in QRS voltage and/or ST segment abnormalities in 6 cases. Echocardiography showed an increase in left ventricular diastolic and/or systolic diameters and volumes in 4 cases but without any decrease in fractional shortening and ejection fraction under normal values: in 2 of them abnormalities of diastolic function (E/A mitral Doppler ratio, isovolumic relaxation time and deceleration time) were also recorded. CONCLUSIONS: Our protocol of cyclophosphamide administration did not cause cardiac toxicity by myocardial cell damage, as analyzed by troponin I levels, thus suggesting that myocyte membrane injury is not the first mechanism of it. ECG (i.e. QRS voltages) and echo-Doppler (i.e. E/A ratio) monitoring lead to hypothesize that endothelial injury and interstitial edema with subsequent reduction in left ventricular diastolic compliance may be the first signs of cardiac dysfunction in this clinical setting.


Subject(s)
Antineoplastic Agents, Alkylating/adverse effects , Breast Neoplasms/drug therapy , Cyclophosphamide/adverse effects , Electrocardiography , Heart Diseases/chemically induced , Heart Diseases/diagnosis , Troponin I/blood , Adult , Antineoplastic Agents, Alkylating/administration & dosage , Cyclophosphamide/administration & dosage , Female , Heart Diseases/blood , Heart Diseases/diagnostic imaging , Humans , Middle Aged , Ultrasonography
8.
Int J Oncol ; 17(5): 1015-8, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11029506

ABSTRACT

Galectin-1 has been demonstrated to be a mediator of T-cell apoptosis acting on activated T-cells and, in a selective manner, on different T leukemia cell lines. Here we show that the sensitivity to galectin-1 is associated with repression of the endogenous galectin-1 gene whereas non-sensitive cells express high levels of galectin-1. Repression of galectin-1 gene in sensitive cells is associated with hyper-methylation of the promoter region. Transient treatment of non-expressing cells with the demethylating agent 5-azacytidine led to irreversible demethylation and subsequent reactivation of galectin-1 gene.


Subject(s)
DNA Methylation , Gene Expression Regulation, Leukemic , Hemagglutinins/genetics , Leukemia, T-Cell/pathology , Neoplasm Proteins/genetics , Antimetabolites, Antineoplastic/pharmacology , Azacitidine/pharmacology , DNA (Cytosine-5-)-Methyltransferases/antagonists & inhibitors , DNA (Cytosine-5-)-Methyltransferases/metabolism , DNA Methylation/drug effects , Enzyme Inhibitors/pharmacology , Galectin 1 , Gene Expression Regulation, Leukemic/drug effects , Hemagglutinins/biosynthesis , Humans , Leukemia, T-Cell/genetics , Leukemia, T-Cell/metabolism , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/biosynthesis , Promoter Regions, Genetic , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/metabolism
9.
Infect Control Hosp Epidemiol ; 21(8): 529-31, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10968721

ABSTRACT

Transient plasma human immunodeficiency virus (HIV) copies were detected by nucleic-acid sequence-based amplification during combination antiretroviral prophylaxis in a healthcare worker who reported a percutaneous injury from a stylet and who remained HIV-antibody-negative. An HIV-specific T-helper response, assessed by interleukin-2 production, was observed when tested at 13 months following the exposure.


Subject(s)
HIV Infections/diagnosis , HIV-1/genetics , Needlestick Injuries , Nucleic Acid Amplification Techniques/methods , Occupational Exposure , RNA, Viral/analysis , Adult , Antiviral Agents/therapeutic use , False Positive Reactions , Female , HIV Infections/prevention & control , Humans
10.
J Clin Invest ; 105(9): 1233-41, 2000 May.
Article in English | MEDLINE | ID: mdl-10791998

ABSTRACT

We have found that EGF-R expression is associated with the development of the Schwann cell-derived tumors characteristic of neurofibromatosis type 1 (NF1) and in animal models of this disease. This is surprising, because Schwann cells normally lack EGF-R and respond to ligands other than EGF. Nevertheless, immunoblotting, Northern analysis, and immunohistochemistry revealed that each of 3 malignant peripheral nerve sheath tumor (MPNST) cell lines from NF1 patients expressed the EGF-R, as did 7 of 7 other primary MPNSTs, a non-NF1 MPNST cell line, and the S100(+) cells from each of 9 benign neurofibromas. Furthermore, transformed derivatives of Schwann cells from NF1(-/-) mouse embryos also expressed the EGF-R. All of the cells or cell lines expressing EGF-R responded to EGF by activation of downstream signaling pathways. Thus, EGF-R expression may play an important role in NF1 tumorigenesis and Schwann cell transformation. Consistent with this hypothesis, growth of NF1 MPNST lines and the transformed NF1(-/-) mouse embryo Schwann cells was greatly stimulated by EGF in vitro and could be blocked by agents that antagonize EGF-R function.


Subject(s)
Epidermal Growth Factor/pharmacology , ErbB Receptors/metabolism , Neurofibromatosis 1/metabolism , Proteins/genetics , Animals , Cell Transformation, Neoplastic , Humans , Mice , Mice, Mutant Strains , Neurilemmoma , Neurofibromin 1 , Rats , Tumor Cells, Cultured
11.
J Biol Chem ; 275(11): 7894-901, 2000 Mar 17.
Article in English | MEDLINE | ID: mdl-10713105

ABSTRACT

We have identified a novel human gene encoding a 59-kDa POZ-AT hook-zinc finger protein (PATZ) that interacts with RNF4, a mediator of androgen receptor activity, and acts as a transcriptional repressor. PATZ cDNA was isolated through a two-hybrid interaction screening using the RING finger protein RNF4 as a bait. In vitro and in vivo interaction between RNF4 and PATZ was demonstrated by protein-protein affinity chromatography and coimmunoprecipitation experiments. Such interaction occurred through a small region of PATZ containing an AT-hook DNA binding domain. Immunofluorescence staining and confocal microscopy showed that PATZ localizes in distinct punctate nuclear regions and colocalizes with RNF4. Functional analysis was performed by cotransfection assays: PATZ acted as a transcriptional repressor, whereas its partner RNF4 behaved as a transcriptional activator. When both proteins were overexpressed a strong repression of the basal transcription was observed, indicating that the association of PATZ with RNF4 switches activation to repression. In addition, RNF4 was also found to associate with HMGI(Y), a chromatin-modeling factor containing AT-hook domains.


Subject(s)
Neoplasm Proteins , Nuclear Proteins , Repressor Proteins/metabolism , Zinc Fingers , Amino Acid Sequence , Animals , DNA-Binding Proteins/metabolism , HMGA1a Protein , High Mobility Group Proteins/metabolism , Humans , Kruppel-Like Transcription Factors , Mice , Molecular Sequence Data , Protein Binding , Repressor Proteins/classification , Repressor Proteins/genetics , Sequence Homology, Amino Acid , Transcription Factors/metabolism , Transcription, Genetic , Two-Hybrid System Techniques , Ubiquitin-Protein Ligases
12.
Oncogene ; 19(54): 6306-16, 2000 Dec 14.
Article in English | MEDLINE | ID: mdl-11175345

ABSTRACT

We report here that overexpression of the tuberous sclerosis-1 (TSC1) gene product hamartin results in the inhibition of growth, as well as changes in cell morphology. Growth inhibition was associated with an increase in the endogenous level of the product of the tuberous sclerosis-2 (TSC2) gene, tuberin. As overexpression of tuberin inhibits cell growth, and hamartin is known to bind tuberin, these results suggested that hamartin stabilizes tuberin and this contributes to the inhibition of cell growth. Indeed, transient transfection of TSC1 increased the endogenous level of tuberin, and transient co-transfection of TSC1 with TSC2 resulted in higher tuberin levels. The stabilization was explained by the finding that tuberin is highly ubiquitinated in cells, while the fraction of tuberin that is bound to hamartin is not ubiquitinated. Co-expression of tuberin stabilized hamartin, which is weakly ubiquitinated, in transiently transfected cells. The amino-terminal two-thirds of tuberin was responsible for its ubiquitination and for stabilization of hamartin. A mutant of tuberin from a patient missense mutation of TSC2 was also highly ubiquitinated, and was unable to stabilize hamartin. We conclude that hamartin is a growth inhibitory protein whose biological effect is likely dependent on its interaction with tuberin.


Subject(s)
Proteins/physiology , Repressor Proteins/metabolism , Ubiquitins/metabolism , Animals , COS Cells , Cell Division , Cell Line, Transformed , Cysteine Endopeptidases , Cysteine Proteinase Inhibitors/pharmacology , Fibroblasts/cytology , Fibroblasts/metabolism , Gene Expression , Genes, Tumor Suppressor , Glycoproteins/pharmacology , Multienzyme Complexes/antagonists & inhibitors , Proteasome Endopeptidase Complex , Proteins/genetics , Rats , Repressor Proteins/genetics , Transfection , Tuberous Sclerosis Complex 1 Protein , Tuberous Sclerosis Complex 2 Protein , Tumor Suppressor Proteins
13.
Biochimie ; 81(4): 381-8, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10401673

ABSTRACT

In this review we summarize the available information on the expression of mammalian galectins in normal and transformed cells. From all these studies it is apparent that each cell might express most of galectins; yet, during development or in various differentiation stages or under different physiological or pathological conditions, one or more galectins are preferentially expressed in each cell type. This implies a fine control of gene expression and suggests that such control should be coordinated. Nevertheless, to date very few studies have been performed on the mechanisms responsible for the regulation of galectin genes. We review the current knowledge on galectin promoter function. We believe that this area of galectin research will expand rapidly in the near future.


Subject(s)
Gene Expression Regulation , Hemagglutinins/genetics , Animals , Cell Line , Galectins , Humans , Tissue Distribution , Transcription, Genetic , Tumor Cells, Cultured
14.
Int J Cancer ; 84(1): 39-43, 1999 Feb 19.
Article in English | MEDLINE | ID: mdl-9988230

ABSTRACT

Galectin-1 and galectin-3 are galactoside-binding proteins involved in different steps of tumor progression and potential targets for therapy. We have investigated the expression of these galectins in 38 human bladder transitional-cell carcinomas of different histological grade and clinical stage and in 5 normal urothelium samples. Galectin-1 mRNA levels were highly increased in most high-grade tumors compared with normal bladder or low-grade tumors. Western blot and immuno-histochemical analysis of normal and neoplastic tissues revealed a higher content of galectin-1 in tumors. Galectin-3 mRNA levels were also increased in most tumors compared with normal urothelium, but levels were comparable among tumors of different histological grade.


Subject(s)
Antigens, Differentiation/biosynthesis , Carcinoma, Transitional Cell/metabolism , Hemagglutinins/biosynthesis , Urinary Bladder Neoplasms/metabolism , Adult , Aged , Aged, 80 and over , Antigens, Differentiation/genetics , Blotting, Northern , Blotting, Western , Carcinoma, Transitional Cell/genetics , Carcinoma, Transitional Cell/pathology , Female , Galectin 1 , Galectin 3 , Hemagglutinins/genetics , Humans , Immunohistochemistry , Male , Middle Aged , RNA, Messenger/analysis , Urethra/metabolism , Urinary Bladder/metabolism , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/pathology
15.
Genomics ; 47(2): 258-65, 1998 Jan 15.
Article in English | MEDLINE | ID: mdl-9479498

ABSTRACT

We have isolated a new human RING-finger gene (RNF4) that encodes a 190-amino-acid protein. RNF4, in addition to the carboxyl-terminally located RING-finger motif, contains two putative nuclear localization signals and stretches of acidic amino acids that are similar to the activation domains of some transcription factors. RNF4 was expressed at low levels in all human tissues examined, with the notable exception of very high expression in the testis. The mouse homolog of RNF4 was abundantly expressed in embryonic tissues from the earliest days postgestation and exhibited a ubiquitous pattern of expression as assessed by in situ hybridization. We have mapped RNF4 to 4p16.3, a chromosome region associated with several genetic and neoplastic diseases. RNF4 spans 47 kb, is composed of eight exons, and maps immediately proximal to the anonymous locus D4S183, between the huntingtin (HD) and the fibroblast growth factor receptor 3 (FGFR3) genes.


Subject(s)
Chromosome Mapping , Chromosomes, Human, Pair 4/genetics , DNA-Binding Proteins/genetics , Nuclear Proteins , Transcription Factors , Zinc Fingers/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , DNA-Binding Proteins/analysis , DNA-Binding Proteins/chemistry , Gene Expression Regulation, Developmental , Humans , Mice , Molecular Sequence Data , Organ Specificity/genetics , Ubiquitin-Protein Ligases
16.
FEBS Lett ; 421(2): 152-8, 1998 Jan 09.
Article in English | MEDLINE | ID: mdl-9468297

ABSTRACT

We conducted by bisulfite genomic sequencing a high resolution study of the methylation of the galectin-1 gene in expressing and nonexpressing tissues. We show that: (i) hypomethylation of galectin-1 promoter correlates with expression; (ii) differences in methylation occur in a small region, which include a CpG cluster; (iii) the density of methyl-CpGs rather than site-specific methylation distinguishes the nonexpressing from the expressing alleles; (iv) the modification profiles in nonexpressing tissues are highly heterogeneous; (v) a single CpG within 1300 bp is always methylated both in expressing and nonexpressing tissues; (vi) these features are conserved in rat and mouse.


Subject(s)
DNA Methylation , Hemagglutinins/genetics , Promoter Regions, Genetic , 3T3 Cells , Animals , Base Sequence , Cell Extracts , CpG Islands , DNA, Complementary , DNA-Binding Proteins/metabolism , Galectin 1 , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Nuclear Proteins/metabolism , Rats , Rats, Sprague-Dawley
17.
J Biol Chem ; 272(4): 2136-42, 1997 Jan 24.
Article in English | MEDLINE | ID: mdl-8999914

ABSTRACT

The Raf-1 serine/threonine protein kinase plays a central role in many of the mitogenic signaling pathways regulating cell growth and differentiation. The regulation of Raf-1 is complex, and involves protein-protein interactions as well as changes in the phosphorylation state of Raf-1 that are accompanied by alterations in its electrophoretic mobility. We have previously shown that a 33-kDa COOH-terminal, kinase-inactive fragment of Raf-1 underwent a mobility shift in response to the stimulation of cells with serum or phorbol esters. Here we demonstrate that treatment of NIH 3T3 cells or Sf9 cells with hydrogen peroxide (H2O2) also induces the mobility shift of the kinase-inactive Raf-1 fragment. A series of deletion mutants of the Raf-1 COOH terminus were analyzed, and the region required for the mobility shift was localized to a 78-amino acid fragment (residues 566-643). Metabolic labeling revealed that the slower migrating forms of the 33-kDa and of the smaller fragment contained phosphorus. Mutation of a previously characterized phosphorylation site, serine 621, to alanine prevented the mobility shift as well as phosphate incorporation or Src and Ras-dependent kinase activation in Sf9 cells when this mutation was engineered into the full-length Raf-1. Mutation of 621 to aspartate yielded a protein that existed in both the shifted and unshifted forms, demonstrating that a negative charge at 621 was necessary, but not sufficient, for the mobility shift to occur; however, its full-length form was still resistant to activation in the Sf9 system. Additional mutation of nearby serine 624 to alanine blocked the shift, implicating this residue as the site of the second of a two-step modification process leading to the slower migrating form. Co-expression of the 33-kDa fragment with an activated form of mitogen-activated protein kinase kinase in NIH 3T3 led to the appearance of the shifted form in a serum-independent manner. These results demonstrate that a mitogen-activated protein kinase kinase-induced event involving modification of serines 621 and 624 leads to the mobility shift of Raf-1.


Subject(s)
Protein Serine-Threonine Kinases/chemistry , Proto-Oncogene Proteins/chemistry , Serine , 3T3 Cells , Animals , Electrophoresis, Polyacrylamide Gel , Hydrogen Peroxide/pharmacology , Mice , Phosphorylation , Point Mutation , Proto-Oncogene Proteins c-raf , Tetradecanoylphorbol Acetate/pharmacology , Transfection , Zinc/pharmacology
18.
Mol Cell Biol ; 16(6): 2736-43, 1996 Jun.
Article in English | MEDLINE | ID: mdl-8649381

ABSTRACT

The galectin-1 gene is developmentally regulated gene whose activity is strongly modulated during cell differentiation and transformation. We have previously shown that galectin-1 promoter constructs are highly active when transiently transfected in cells both expressing and not expressing the endogenous gene and that the basal activity is determined by a small region encompassing the transcription start site (from positions -50 to +50). We have now investigated the role of DNA methylation in galectin-1 gene expression. Southern blot analysis with HpaII and MspI endonucleases and sodium bisulfite analysis of genomic DNA from expressing and nonexpressing cell lines and cell hybrids showed a close correlation between gene activity and demethylation of the 5' region of the galectin-1 gene. We found that the galectin-1 promoter region is fully methylated, at every CpG site on both strands, in nonexpressing differentiated rat liver (FAO) and thyroid (PC C13) cells and unmethylated in the expressing undifferentiated liver (BRL3A) and thyroid transformed (PC myc/raf) cell lines. In addition, reactivation of the silent FAO alleles in FAO-human osteosarcoma (143tk-) hybrid cells is accompanied by a complete demethylation of the promoter region. Finally, when galectin-1 chloramphenicol acetyltransferase (CAT) promoter constructs were methylated in vitro by SssI methylase at every cytosine residue of the CpG doublets and transfected into mouse fibroblasts, the transcription of the CAT reporter gene was strongly inhibited.


Subject(s)
DNA/genetics , DNA/metabolism , Gene Expression Regulation, Developmental , Hemagglutinins/genetics , Lectins/genetics , Animals , Base Sequence , Cell Line , CpG Islands , DNA Primers/genetics , Deoxyribonuclease HpaII , Galectin 1 , Humans , Hybrid Cells , Methylation , Mice , Molecular Sequence Data , Promoter Regions, Genetic , Rats , Sulfites
19.
FEBS Lett ; 373(2): 159-63, 1995 Oct 09.
Article in English | MEDLINE | ID: mdl-7589457

ABSTRACT

The galectin-1 gene encodes a beta-galactoside-binding protein whose overexpression is associated with neoplastic transformation and loss of differentiation. Transient transfection assays of a series of deletions constructs (pGAT) showed that the galectin-1 promoter is highly active in cells both expressing and non-expressing the endogenous gene, and that the basal activity is determined by sequences encompassing the transcription start site (-50/+50). Both an upstream (-50/-26) and a downstream position-dependent (+10/+50) cis-elements are necessary for efficient transcriptional activity and are able to bind nuclear proteins.


Subject(s)
Hemagglutinins/biosynthesis , Hemagglutinins/genetics , Promoter Regions, Genetic , 3T3 Cells , Animals , Base Sequence , Cell Differentiation , Cell Line , Cell Transformation, Neoplastic , Chloramphenicol O-Acetyltransferase/biosynthesis , Galectin 1 , Gene Expression , Lectins/biosynthesis , Lectins/genetics , Mice , Molecular Sequence Data , Plasmids , Rats , Recombinant Proteins/biosynthesis , Restriction Mapping , Sequence Deletion , Transfection
20.
Int J Cancer ; 64(3): 171-5, 1995 Jun 22.
Article in English | MEDLINE | ID: mdl-7622305

ABSTRACT

We previously reported that galectin-1 gene expression increases up to 100-fold in oncogene-transformed rat thyroid cells compared with their normal counterparts and that the relative mRNA levels correlate with the degree of malignancy. In the present study we investigated whether galectin-1 is differentially expressed in human thyroid neoplasms, which range from well-differentiated tumors to undifferentiated anaplastic carcinomas. We analyzed 74 human thyroid specimens of neoplastic, hyperproliferative and normal tissues and several tumor cell lines. Galectin-1 mRNA and protein levels were higher in 6 thyroid carcinoma-derived cell lines than in normal thyroid primary cultures and adenoma cells. Galectin-1 mRNA levels increased in 28/40 papillary carcinomas and in 6/7 anaplastic carcinomas compared with normal or hyperplastic thyroid. Conversely, galectin-1 expression was unaffected in follicular carcinomas and benign adenomas. Immunohistochemical analysis of normal thyroid and papillary carcinoma sections revealed a higher content of galectin-1 protein in neoplastic follicular cells than in normal cells.


Subject(s)
Hemagglutinins/analysis , Lectins/analysis , Thyroid Gland/chemistry , Thyroid Neoplasms/chemistry , Base Sequence , Galectin 1 , Humans , Immunohistochemistry , Molecular Sequence Data , Thyroid Neoplasms/pathology , Tumor Cells, Cultured
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