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1.
Cell Death Dis ; 6: e1909, 2015 Oct 15.
Article in English | MEDLINE | ID: mdl-26469958

ABSTRACT

The mevalonate (MVA) pathway is an important metabolic pathway implicated in multiple aspects of tumorigenesis. In this study, we provided evidence that p53 induces the expression of a group of enzymes of the MVA pathway including 3'-hydroxy-3'-methylglutaryl-coenzyme A reductase, MVA kinase, farnesyl diphosphate synthase and farnesyl diphosphate farnesyl transferase 1, in the human glioblastoma multiforme cell line, U343 cells, and in normal human astrocytes, NHAs. Genetic and pharmacologic perturbation of p53 directly influences the expression of these genes. Furthermore, p53 is recruited to the gene promoters in designated p53-responsive elements, thereby increasing their transcription. Such effect was abolished by site-directed mutagenesis in the p53-responsive element of promoter of the genes. These findings highlight another aspect of p53 functions unrelated to tumor suppression and suggest p53 as a novel regulator of the MVA pathway providing insight into the role of this pathway in cancer progression.


Subject(s)
Brain Neoplasms/metabolism , Glioblastoma/metabolism , Mevalonic Acid/metabolism , Tumor Suppressor Protein p53/physiology , Cell Line, Tumor , Cholesterol/biosynthesis , Gene Expression Regulation, Neoplastic , Humans , Metabolic Networks and Pathways , Promoter Regions, Genetic , Transcription, Genetic
2.
Br J Pharmacol ; 159(4): 831-41, 2010 Feb.
Article in English | MEDLINE | ID: mdl-20067470

ABSTRACT

BACKGROUND AND PURPOSE: The endocannabinoid system and the cannabinoid CB(1) receptor have been identified in human sperm, and it is well known that endocannabinoids have pronounced adverse effects on male and female reproduction. In order to elucidate further the pathophysiological role of the endocannabinoid system in male fertility, we investigated the activity of the CB(1) receptor antagonist rimonabant (SR141716) on the fertilizing ability of human sperm. EXPERIMENTAL APPROACH: We evaluated in vitro the effects of rimonabant on motility, survival, capacitation, acrosin activity and metabolism of human sperm. Particularly, capacitation was studied by using three different approaches: intracellular free Ca(2+) content assay, cholesterol efflux assay and protein tyrosine phosphorylation analysis. KEY RESULTS: Rimonabant significantly increased sperm motility and viability through the induction of pAkt and pBcl2, key proteins of cell survival and metabolism, and it induced acrosome reaction and capacitation as well. Rimonabant reduced the triglyceride content of sperm, while enhancing lipase and acyl-CoA dehydrogenase activities, implying an overall lipolytic action in these cells. Rimonabant also affected sperm glucose metabolism by decreasing phosphorylation of glycogen synthase kinase 3 and increasing glucose-6-phosphate dehydrogenase activity, suggesting a role in inducing sperm energy expenditure. Intriguingly, agonism at the CB(1) receptor, with an anandamide analogue or a selective inhibitor of fatty acid amide hydrolase, produced opposing effects on human sperm functions. CONCLUSIONS AND IMPLICATIONS: Our data suggest that blockade of the CB(1) receptor by rimonabant induces the acquisition of fertilizing ability and stimulates energy expenditure in human sperm.


Subject(s)
Energy Metabolism/drug effects , Fertilization/drug effects , Piperidines/pharmacology , Pyrazoles/pharmacology , Receptor, Cannabinoid, CB1/antagonists & inhibitors , Spermatozoa/drug effects , Acrosin/metabolism , Acrosome Reaction/drug effects , Acyl-CoA Dehydrogenase/metabolism , Arachidonic Acids/pharmacology , Calcium/metabolism , Cell Survival/drug effects , Cholesterol/metabolism , Dose-Response Relationship, Drug , Endocannabinoids , Glucose/metabolism , Glucosephosphate Dehydrogenase/metabolism , Glycogen Synthase Kinase 3/metabolism , Humans , Lipase/metabolism , Male , Phosphorylation , Polyunsaturated Alkamides/pharmacology , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Receptor, Cannabinoid, CB1/agonists , Receptor, Cannabinoid, CB1/metabolism , Rimonabant , Sperm Capacitation/drug effects , Sperm Motility/drug effects , Spermatozoa/metabolism , Triglycerides/metabolism , Tyrosine
3.
Genes Brain Behav ; 8(7): 728-32, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19659925

ABSTRACT

Endocannabinoids modulate eating behavior; hence, endocannabinoid genes may contribute to the biological vulnerability to eating disorders. The rs1049353 (1359 G/A) single nucleotide polymorphism (SNP) of the gene coding the endocannabinoid CB1 receptor (CNR1) and the rs324420 (cDNA 385C to A) SNP of the gene coding fatty acid amide hydrolase (FAAH), the major degrading enzyme of endocannabinoids, have been suggested to have functional effects on mature proteins. Therefore, we explored the possibility that those SNPs were associated to anorexia nervosa and/or bulimia nervosa. The distributions of the CNR1 1359 G/A SNP and of the FAAH cDNA 385C to A SNP were investigated in 134 patients with anorexia nervosa, 180 patients with bulimia nervosa and 148 normal weight healthy controls. Additive effects of the two SNPs in the genetic susceptibility to anorexia nervosa and bulimia nervosa were also tested. As compared to healthy controls, anorexic and bulimic patients showed significantly higher frequencies of the AG genotype and the A allele of the CNR1 1359 G/A SNP. Similarly, the AC genotype and the A allele of the FAAH cDNA 385C to A SNP were significantly more frequent in anorexic and bulimic individuals. A synergistic effect of the two SNPs was evident in anorexia nervosa but not in bulimia nervosa. Present findings show for the first time that the CNR1 1359 G/A SNP and the FAAH cDNA 385C to A SNP are significantly associated to anorexia nervosa and bulimia nervosa, and demonstrate a synergistic effect of the two SNPs in anorexia nervosa.


Subject(s)
Amidohydrolases/genetics , Anorexia Nervosa/genetics , Bulimia Nervosa/genetics , Cannabinoid Receptor Modulators/metabolism , Endocannabinoids , Polymorphism, Single Nucleotide/genetics , Receptor, Cannabinoid, CB1/genetics , Adult , Anorexia Nervosa/metabolism , Anorexia Nervosa/physiopathology , Brain/metabolism , Brain/physiopathology , Brain Chemistry/genetics , Bulimia Nervosa/metabolism , Bulimia Nervosa/physiopathology , DNA Mutational Analysis , Energy Metabolism/genetics , Female , Gene Frequency/genetics , Genetic Markers , Genetic Predisposition to Disease/genetics , Genetic Testing , Genotype , Humans , Male , Phenotype , Young Adult
4.
Br J Pharmacol ; 153(5): 1003-10, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18157162

ABSTRACT

BACKGROUND AND PURPOSE: Rimonabant (SR141716) is the first selective cannabinoid receptor CB(1) antagonist described. Along with its anti-obesity action, emerging findings show potential anti-proliferative and anti-inflammatory action of SR141716 in several in vitro and in vivo models. In this study we have investigated the anti-proliferative and immunomodulatory effects of SR141716 in human peripheral blood mononuclear cells (PBMCs). EXPERIMENTAL APPROACH: We have evaluated in vitro the effect of SR141716 in human PBMCs stimulated with different mitogens. Cell proliferation was assessed by (3)H-thymidine incorporation. Cell cycle, cell death and apoptosis were analysed by flow cytometry. Protein expression was investigated by Western blot. KEY RESULTS: SR141716 significantly inhibited the proliferative response of PBMCs and this effect was accompanied by block of G(1)/S phase of the cell cycle without induction of apoptosis and cell death. SR141716 used in combination with 2-methyl-arachidonyl-2'-fluoro-ethylamide (Met-F-AEA), a stable analogue of the endogenous cannabinoid anandamide, showed synergism rather than antagonism of the inhibition of cell proliferation. The immunomodulatory effects of SR141716 were associated with increased expression of IkappaB, phosphorylated AKT (p-AKT) and decreased expression of NF-kappaB, p-IkappaB, p-ERK, COX-2 and iNOS. CONCLUSIONS AND IMPLICATIONS: Our findings suggest SR141716 is a novel immunomodulatory drug with anti-inflammatory properties.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Leukocytes, Mononuclear/drug effects , Piperidines/pharmacology , Pyrazoles/pharmacology , Receptor, Cannabinoid, CB1/antagonists & inhibitors , Apoptosis/drug effects , Arachidonic Acids/pharmacology , Cell Proliferation/drug effects , Cells, Cultured , Drug Synergism , G1 Phase/drug effects , Gene Expression Regulation/drug effects , Humans , I-kappa B Proteins/drug effects , I-kappa B Proteins/metabolism , Leukocytes, Mononuclear/metabolism , Phosphorylation/drug effects , Proto-Oncogene Proteins c-akt/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Rimonabant , S Phase/drug effects
5.
Int J Obes (Lond) ; 31(6): 908-12, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17160086

ABSTRACT

CONTEXT: Endocannabinoids control food intake via both central and peripheral mechanisms, and cannabinoid type-1 receptor (CB1) modulates lipogenesis in primary adipocyte cell cultures and in animal models of obesity. OBJECTIVES: We aimed to evaluate, at the population level, the frequency of a genetic polymorphism of CB1 and to study its correlation with body mass index. DESIGN, SETTING AND PARTICIPANTS: Healthy subjects from a population survey carried out in southern Italy examined in 1992-1993 and older than 65 years (n=419, M=237, F=182) were divided into quintiles by body mass index (BMI). Two hundred and ten subjects were randomly sampled from the first, third and fifth quintile of BMI (BMI, respectively: 16.2-23.8=normal, 26.7-28.4=overweight, 31.6-49.7=obese) to reach a total of 70 per quintile. Their serum and white cells from the biological bank were used to measure the genotype and the blood variables for the study. MEASUREMENTS: Anthropometric parameters, blood pressure, serum glucose and lipid levels were measured with standard methods; genotyping for the CB1 1359G/A polymorphism was performed using multiplex PCR. Statistical methods included chi2 for trend, binomial and multinomial multiple logistic regression to model BMI on the genotype, controlling for potential confounders. RESULTS: We found a clear trend of increasing relative frequency of the CB1 wild-type genotype with the increase of BMI (P=0.03) and, using a multiple logistic regression model, wild-type genotype, female gender, age, glycaemia and triglycerides were directly associated with both overweight (third quintile of BMI) and obesity (fifth quintile of BMI). CONCLUSIONS: Although performed in a limited number of subjects, our results show that the presence of the CB1 polymorphic allele was significantly associated with a lower BMI.


Subject(s)
Body Mass Index , Polymorphism, Genetic/genetics , Receptor, Cannabinoid, CB1/genetics , Age Distribution , Aged , Blood Glucose/analysis , Female , Genotype , Humans , Italy/epidemiology , Male , Population Surveillance/methods , Regression Analysis , Sex Distribution , Triglycerides/blood
6.
J Viral Hepat ; 10(4): 266-70, 2003 Jul.
Article in English | MEDLINE | ID: mdl-12823592

ABSTRACT

Mutated p53 acts as a dominant oncogene and alterations in the p53 gene are described in a large number of patients with hepatocellular carcinoma (HCC). It has been demonstrated that hepatitis C virus (HCV)-core protein regulates transcriptionally cellular genes, as well as cell growth and apoptosis. This study was undertaken to evaluate whether p53 may be expressed also in a precocious stage of HCV-related liver damage. We studied p53 expression by immunoluminometric assay on liver samples from 40 patients (M/F 18/ 22, median age 44 years, range 13-64 years) with biopsy-proven HCV-related chronic hepatitis. We considered the following factors: degree of liver damage, liver iron content and HCV-RNA titre. We also evaluated as possible co-factors alcohol and food intake in the last 3 years. p53 was over-expressed in seven of 40 (17.5%) patients. Liver histology documented the presence of unexpected cirrhosis in two patients among the p53 positive subjects. The p53 positive group had a daily ethanol intake significantly higher in respect to that of the p53 negative group (P < 0.05). Alimentary history documented that patients with a p53 over-expression had a lower intake of total calories, monounsaturated fatty acids, vitamin C and riboflavin. Data indicate that p53 over-expression can occur even in initial stages of HCV-related liver disease.


Subject(s)
Genetic Predisposition to Disease , Hepacivirus/genetics , Hepatitis C, Chronic/genetics , Hepatitis C, Chronic/pathology , Tumor Suppressor Protein p53/genetics , Adolescent , Adult , Analysis of Variance , Biopsy, Needle , Case-Control Studies , Chi-Square Distribution , Cohort Studies , DNA, Viral/analysis , Female , Gene Expression Regulation, Viral , Humans , Immunohistochemistry , Male , Middle Aged , Probability , Proto-Oncogene Proteins/genetics , Reference Values , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity , Statistics, Nonparametric
7.
Ann N Y Acad Sci ; 963: 329-31, 2002 Jun.
Article in English | MEDLINE | ID: mdl-12095960

ABSTRACT

Helicobacter pylori is a definite carcinogen whose mechanism of action is still unknown. The aim of this work was (1) to determine the presence of p53 protein and related antibodies in patients affected by various gastric pathologies and chronically infected with H. pylori, and (2) to try to discover a test to be used as a marker of a possible switch towards a neoplastic phenotype.


Subject(s)
Cell Transformation, Neoplastic/metabolism , Helicobacter Infections/physiopathology , Helicobacter pylori , Tumor Suppressor Protein p53/metabolism , Antibodies/immunology , Cell Transformation, Neoplastic/immunology , Helicobacter Infections/immunology , Helicobacter Infections/metabolism , Humans , Phenotype , Tumor Suppressor Protein p53/immunology
8.
Mol Med ; 7(8): 552-60, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11591891

ABSTRACT

BACKGROUND: The retinoblastoma-interacting zinc-finger gene RIZ is expressed in two forms (RIZ1 and RIZ2) that differ for the presence near the N-terminus of RIZ1 of a conserved domain, defined PR (PRDI-BF1-RIZ homology), homologous to a similar domain present in other proteins recognized as tumor suppressor gene products. The RIZ1 form is usually absent or expressed at low levels in tumor cells, whereas RIZ2 is frequently expressed. We investigated a possible involvement of RIZ1 in differentiation control using a myeloid cell maturation model that is easily modulated by retinoids and other agents. MATERIALS AND METHODS: HL60 or NB4 cell lines or patients' leukemic promyelocytes were treated with all- trans -retinoic acid or other agents to induce differentiation. RIZ gene expression was determined with reverse transcriptase polymerase chain reaction (RT-PCR) and RNase protection assay. Immunocytochemistry was performed to assess variation of the intracellular distribution of RIZ protein on all- trans-retinoic acid treatment. Forced expression of RIZ1 protein was obtained with a recombinant adenovirus containing RIZ1 cDNA. RESULTS: Treatment with retinoic acid induced a selective expression of RIZ1 in HL60 cell line. Retinoic acid effect was maximal at 7 days and correlated to the granulocytic differentiation of cells. A similar effect was obtained in retinoic acid-sensitive NB4 cell line or in patients' leukemic promyelocytes, but not in the retinoic acid-resistant cell line NB4.007/6 or in the U937 cell line. Selective expression of RIZ1 was also induced by 12-O-tetradecanoyl-phorbol-13-acetate in the U937 and HL60 cell lines and by 1,25-dihydroxyvitamin D(3) only in HL60 cells. In HL60 cells, RIZ1 was also induced by activation of a retinoid alpha receptor-independent maturation pathway based on retinoid X receptor agonist and protein kinase A synergism. In addition, retinoic acid produced a redistribution of the antigen within the nucleus in these cells. Forced expression of RIZ1 protein induced growth arrest and death of HL60 cells. CONCLUSIONS: The correlation between the selective expression of RIZ1 induced by retinoic acid, 12-O-tetradecanoyl-phorbol-13-acetate, or 1,25-dihydroxyvitamin D(3) and differentiation suggested that RIZ protein was involved in myeloid cell differentiation induced by these agents.


Subject(s)
Cell Differentiation/physiology , DNA-Binding Proteins , Myeloid Cells/physiology , Nuclear Proteins/metabolism , Transcription Factors , Adenosine Monophosphate/analogs & derivatives , Adenoviridae/metabolism , Antineoplastic Agents/pharmacology , Benzoates/pharmacology , Cells, Cultured , Histone-Lysine N-Methyltransferase , Humans , Immunoblotting , Immunohistochemistry , Myeloid Cells/cytology , Myeloid Cells/drug effects , Nuclear Proteins/genetics , Retinoblastoma Protein/genetics , Retinoblastoma Protein/metabolism , Retinoids/pharmacology , Tretinoin/pharmacology , Zinc Fingers/genetics
9.
Lung Cancer ; 33(2-3): 163-9, 2001.
Article in English | MEDLINE | ID: mdl-11551411

ABSTRACT

As reported earlier, p53 antibodies are detected in the sera of patients with different types of cancer, including lung cancer. In contrast, in the serum of healthy subjects the presence of anti-p53 antibodies is extremely rare. We collected the venous blood samples of 109 patients affected with lung cancer (LC): 57 patients (46 M, 11 F) with non-small-cell carcinoma (NSCLC), 52 others (40 M, 12 F) with small-cell carcinoma (SCLC). Serum p53 antibodies were assayed using ELISA method and all positive sera were confirmed by Western-blot method. In addition, using IRMA methods we assayed serum CEA, TPA, CYFRA21-1 and NSE. Serum p53Ab are detectable (p53Ab-positive) in 35/109 (32.1%) patients with lung cancer. About 17/57 (29.8%) patients affected with NSCLC and 18/52 (34.6%) with SCLC were p53Ab-positive. CEA, TPA, CYFRA21-1 and NSE sensitivity in LC patients (NSCLC+SCLC) is 50.5%, 58.7%, 42.2%, 35.8%, respectively. The lower sensitivity (32.1%) of serum p53Ab is connected with the higher specificity and diagnostic accuracy (100% and 69%, respectively). Out of 35 patients p53Ab-positive, five (14.3%) exhibit only serum p53Ab, while serum values of the established tumor markers were lower than cut-off. Serum p53Ab assessment is a simple and a low-cost assay with a good specificity and diagnostic accuracy that in LC patients can be used at least in association with established tumor markers.


Subject(s)
Antibodies, Neoplasm/blood , Antigens, Neoplasm/immunology , Biomarkers, Tumor/blood , Carcinoma, Non-Small-Cell Lung/immunology , Carcinoma, Small Cell/immunology , Lung Neoplasms/immunology , Tumor Suppressor Protein p53/immunology , Aged , Aged, 80 and over , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Neoplasm Staging , Sensitivity and Specificity
10.
J Pediatr Endocrinol Metab ; 14(9): 1635-9, 2001.
Article in English | MEDLINE | ID: mdl-11795654

ABSTRACT

OBJECTIVE: It is rare that manufacturers report age-related FT3, FT4 and TSH normal ranges in healthy children. STUDY DESIGN: Using a solid phase time-resolved fluoroimmunometric method, we determined serum FT3, FT4 and TSH in 3,360 healthy children, age 2-16 years, that we grouped into two age ranges (2-7 yr and 9-16 yr). RESULTS: Boys' and girls' mean serum thyroid hormone values substantially overlap in all age groups. In the age range 2-7 yr, FT3, FT4 and TSH values were 0.10-0.67 ng/dl (mean 0.37 ng/dl), range 0.45-2.29 ng/dl (mean 1.18 ng/dl) and 0.10-5.9 microU/ml (mean 2.2 microU/ml), respectively. In the age range 9-16 yr, FT3, FT4 and TSH values were 0.11-0.53 ng/dl (mean 0.35 ng/dl), 0.69-1.69 ng/dl (mean 1.07 ng/dl) and 0.20-6.1 microU/ml (mean 2.3 pU/ml), respectively. CONCLUSION: Our results represent a useful set of reference values in children and can help physicians in the management of thyroid diseases.


Subject(s)
Thyrotropin/blood , Thyroxine/blood , Triiodothyronine/blood , Adolescent , Aging/blood , Child , Child, Preschool , Female , Humans , Male , Osmolar Concentration , Reference Values
13.
Tumori ; 85(1): 32-4, 1999.
Article in English | MEDLINE | ID: mdl-10228494

ABSTRACT

AIMS AND BACKGROUND: E-cadherin, also known as uvomorulin or cell-CAM 120/80, is one of the subclasses of cadherins, CA(2+)-dependent cell adhesion molecules. Several recent studies have suggested that loss of E-cadherin may be associated with tumor progression, such as in lung, gastric, hepatocellular, breast and prostatic carcinoma. Assessment of E-cadherin serum levels in lung cancer showed a relation to histologic type. METHODS AND STUDY DESIGN: Using an enzyme immunoassay, we determined E-cadherin serum levels in 79 patients affected with lung cancer (stage I-IV), 9 patients with breast cancer, 23 patients with different benign diseases, and 20 healthy patients. RESULTS: At a specificity level of 90%, E-cadherin diagnostic sensitivity was 66.6%, 47.6% and 43.7% in patients affected with squamous cell carcinoma, small cell carcinoma and adenocarcinoma, respectively. CONCLUSIONS: Preliminary results suggest the use of serum E-cadherin as a prospective tumor marker.


Subject(s)
Cadherins/blood , Carcinoma, Non-Small-Cell Lung/blood , Carcinoma, Small Cell/blood , Lung Neoplasms/blood , Lung Neoplasms/pathology , Carcinoma, Non-Small-Cell Lung/pathology , Carcinoma, Small Cell/pathology , Female , Humans , Male , Neoplasm Staging , Sensitivity and Specificity
14.
Tumori ; 83(2): 594-8, 1997.
Article in English | MEDLINE | ID: mdl-9226027

ABSTRACT

An immunoradiometric method of the second generation (IR-MA II) is widely used to determine CA125 serum levels. In this study we have evaluated the performance characteristics of a commercially available IRMA CA125 II (Byk-Gulden, Sangtec Diagnostica). The CA125 serum levels were determined in several groups of patients (healthy women, pregnant women, subjects affected by benign and malignant ovarian cancer, patients with liver diseases) with two IRMAs CA125 II (Byk-Gulden, Sangtec Diagnostica and Centocor, Diagnostic Division) and IRMA CA125 I (Byk-Gulden, Sangtec Diagnostica). Our results show a good analytic performance of IRMA CA125 II (Byk-Gulden, Sangtec Diagnostica), a good correlation between IRMAs CA125 II (Byk-Gulden, Sangtec Diagnostica and Centocor, Diagnostic Division), but an unacceptable correlation between IRMAs CA125 II (Byk-Gulden, Sangtec Diagnostica and Centocor, Diagnostic Division) and IRMA CA125 I. A statistically significant difference was observed comparing the values obtained with both IRMAs CA125 II and IRMA CA125 I in the groups of patients. In contrast no statistically significant difference was observed when we compared the values obtained with IRMA CA125 II (Byk-Gulden, Sangtec Diagnostica) and IRMA CA125 II (Centocor, Diagnostic Division). CA125 serum values obtained with the second-generation kits were different from those obtained with the first-generation one; consequently, it is important, especially in the follow-up of cancer patients, that CA125 serum values be obtained with kits of the same generation. Our data seem to suggest the use of second-generation kits to determine CA125 serum levels.


Subject(s)
CA-125 Antigen/blood , Immunoradiometric Assay , Liver Diseases/immunology , Ovarian Diseases/immunology , Ovarian Neoplasms/immunology , Pregnancy/immunology , Female , Humans , Reference Values
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