29 research outputs found
Use of FTA® classic cards for epigenetic analysis of sperm DNA
No full textFTA® technologies provide the most reliable method for DNA extraction. Although FTA technologies have been widely used for genetic analysis, there is no literature on their use for epigenetic analysis yet. We present for the first time, a simple method for quantitative methylation assessment based on sperm cells stored on Whatman FTA classic cards. Specifically, elution of seminal DNA from FTA classic cards was successfully tested with an elution buffer and an incubation step in a thermocycler. The eluted DNA was bisulfite converted, amplified by PCR, and a region of interest was pyrosequenced
Spartin: At the crossroad between ubiquitination and metabolism in cancer
No full textSPART is a gene coding for a multifunctional protein called spartin, localized in various organelles of human cells. Mutations in the coding region are responsible for a hereditary form of spastic paraplegia called Troyer syndrome while the epigenetic silencing has been demonstrated for some types of tumors. The main functions of this gene are associated to endosomic trafficking and receptor degradation, microtubule interaction, cytokinesis, fatty acids and oxidative metabolism. Spartin has been shown to be a target regulated by STAT3 and localizes also at the level of the mitochondrial outer membrane, where it forms part of a complex maintaining the integrity of the membrane potential. The most recent evidences report a downregulation of spartin in tumor tissues when compared to adjacent normal samples. This intriguing evidence supports further research aimed at clarifying the role of this protein in cancer development and metabolism
Droplet digital PCR is a sensitive tool for the detection of TP53 deletions and point mutations in chronic lymphocytic leukaemia
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Integrative systems medicine approaches to identify molecular targets in lymphoid malignancies
No full textAlthough survival rates for lymphoproliferative disorders are steadily increasing both in the US and in Europe, there is need for optimizing front-line therapies and developing more effective salvage strategies. Recent advances in molecular genetics have highlighted the biological diversity of lymphoproliferative disorders. In particular, integrative approaches including whole genome sequencing, whole exome sequencing, and transcriptome or RNA sequencing have been instrumental to the identification of molecular targets for treatment. Herein, we will discuss how genomic, epigenomic and proteomic approaches in lymphoproliferative disorders have supported the discovery of molecular lesions and their therapeutic targeting in the clinic
Abstract 5: The effects of SIRT1 inhibitors nicotinamide and Ex-527 on lymphoma cells
No full textAbstract
SIRT1 is a well-known lysine-deacetylase, having both histone and non-histone proteins as molecular targets. SIRT1 may function as a tumor promoter as well as a tumor suppressor in a context-dependent manner. We previously demonstrated that the treatment with resveratrol at concentrations able to induce apoptosis leads to SIRT1 down-regulation in lymphoma cells. Here we look for SIRT1 inhibition in order to investigate the effects on proliferation potential, apoptosis induction and targets modulation at a molecular level. The aim is to study whether SIRT1 inhibition affects cell cycle and proliferation capability of lymphoma cells. Experimental procedures. Diffuse large-B cell lymphoma derived cell line Toledo; human lymphoblastoid cell line GGB #7; SIRT1 inhibitors: nicotinamide (NAM) and Ex-527; cell cycle analysis; caspase-3 activity fluorimetric assay; genotoxicity assay gamma-H2AX; fluorescence-activated cell sorting intracellular staining for acetylated-H4K16 histone. New data. We found that the calculated IC50s for SIRT1 inhibition is 50mM for NAM and 343μM for Ex-527. The treatment with either NAM or Ex-527 leads to cell cycle arrest in both Toledo and GGB#7 cells, although in a different fashion. Namely, NAM causes an S-phase accumulation of Toledo while it leads GGB #7 to G0/G1 phase arrest with a concomitant decrease in S-phase already after 10mM. In both cell lines NAM 50mM induces apoptosis as demonstrated by caspase-3 activation and sub-G1 peak appearance in the cell cycle profiles. Ex-527 causes an accumulation in the S phase of Toledo cells and a decrease of G2/M in GGB#7 cells. No induction of apoptosis at the IC50 concentration was observed in both cell lines. Genotoxicity of the above mentioned treatments was investigated through the quantitation of gamma-H2AX histone released by treated cells. No genotoxic effects have been observed upon the treatments with these two inhibitors even at the concentration where NAM induces apoptosis. In order to correlate the observed changes with SIRT1 inhibition, we studied the acetylated H4K16 histone, a direct target of SIRT1 deacetylation. Our preliminary data indicate that NAM treatment increases the percentage of acetylated H4K16. Conclusions. Lymphoma and lymphoblastoid cells are affected by SIRT1 inhibition. NAM and Ex-527 cause cell cycle arrest and growth inhibition. Only NAM 50mM induces apoptosis without genotoxic effects. Overall, our data suggest the direct involvement of SIRT1 during the observed cell cycle arrest and growth inhibition mediated by these inhibitors.
Citation Format: Manuela Guardi, Mariaelena Pistoni, Raffaele Frazzi. The effects of SIRT1 inhibitors nicotinamide and Ex-527 on lymphoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5. doi:10.1158/1538-7445.AM2017-5</jats:p
Study on the role of clusterin in Hodgkin lymphoma
No full textClusterina è una proteina da stress ampiamente espressa nei tessuti e nei fluidi umani. Tuttavia vi sono poche informazioni riguardo alla sua funzione nei linfomi. In questo lavoro dimostriamo che la sua espressione in linee cellulari di linfoma di Hodgkin aumenta in risposta a diversi stimoli tra cui doxorubicina, interferon-gamma ed alte dosi di raggi X. La sua forma circolante è dosabile nel siero di pazienti affetti da linfoma di Hodgkin e sembra essere correlata con la persistenza del tessuto neoplastico ipercaptante dopo due cicli di chemioterapia.Clusterin is a molecular chaperon widely studied in cancers of epithelial origin. On the contrary, the information available on its functions in lymphomas are still quite poor. Here we demonstrate clusterin up-regulation in Hodgkin lymphoma derived cell lines as caused by doxorubicin, interferon-gamma and high doses of X-rays. We also present preliminary data on the quantitation of secretory clusterin in sera from Hodgkin lymphoma patients. The preliminary data show a correlation between the persistence of hypercaptating neoplastic mass and the increase in circulating clusterin
SIRT1 in Secretory Organ Cancer
No full textMammalian silent information regulator 1 (SIRT1) is reported to play a role in cancers of the secretory organs, including thyroid, pancreatic endocrine, and ovarian tumors [1, 2, 3, 4]. A recent meta-analysis conducted on 37 selected studies of human cancers analyzed the correlations of overall survival (OS), disease-free survival (DFS) and relapse-free survival (RFS) with SIRT1 expression [5]. This study reported that SIRT1 overexpression was associated with a worse OS in liver and lung cancers, while it was not correlated with OS in breast cancer, colorectal cancer, or gastric carcinoma. Collectively, the meta-analysis revealed that an unfavorable OS was associated with SIRT1 expression for solid malignancies. Given the growing importance of this class of lysine/histone deacetylases in human endocrine malignancies, a rational and focused literature assessment is desirable in light of future clinical translations
Study on the role of clusterin in Hodgkin lymphoma
No full textClusterina è una proteina da stress ampiamente espressa nei tessuti e nei fluidi umani. Tuttavia vi sono poche informazioni riguardo alla sua funzione nei linfomi. In questo lavoro dimostriamo che la sua espressione in linee cellulari di linfoma di Hodgkin aumenta in risposta a diversi stimoli tra cui doxorubicina, interferon-gamma ed alte dosi di raggi X. La sua forma circolante è dosabile nel siero di pazienti affetti da linfoma di Hodgkin e sembra essere correlata con la persistenza del tessuto neoplastico ipercaptante dopo due cicli di chemioterapia.Clusterin is a molecular chaperon widely studied in cancers of epithelial origin. On the contrary, the information available on its functions in lymphomas are still quite poor. Here we demonstrate clusterin up-regulation in Hodgkin lymphoma derived cell lines as caused by doxorubicin, interferon-gamma and high doses of X-rays. We also present preliminary data on the quantitation of secretory clusterin in sera from Hodgkin lymphoma patients. The preliminary data show a correlation between the persistence of hypercaptating neoplastic mass and the increase in circulating clusterin.ope
The Multiple Mechanisms of Cell Death Triggered by Resveratrol in Lymphoma and Leukemia
No full textLymphoma and leukemia represent a serious threat to human health and life expectancy. Resveratrol is, among the natural-derived chemopreventive molecules, one of the most effective and better studied. In this paper the main mechanisms of cell death triggered by- or linked to- resveratrol are reviewed and discussed. The main focus is on lymphoma and leukemia experimental models where resveratrol has been tested and investigated at the cellular, molecular or physiological levels. The most relevant in vivo challenges involving resveratrol are also reported and analyzed in order to define the key features of this polyphenol and the potential for the treatment of hematologic tumors
Plasmid DNA And IL-4 Modulate Expression of MHC Class I And Costimulatory Molecules in B Lymphocytes
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