Papers by Federica Barbieri
In this work high resolution techniques as transmission electron microscopy (TEM), scanning elect... more In this work high resolution techniques as transmission electron microscopy (TEM), scanning electron microscopy (SEM), Zeta Potential measurements, and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF) have been employed to deeply investigate about silver nanoparticles (AgNPs) electrochemically synthesized and successfully applied in biological and chemical fields. Strong brightness, as well as the tendency to generate odd number nanoclusters, and the absence of free silver ion in solution have been observed. The chemical and physical properties of the AgNPs seem to be related to their peculiar oxidative state as suggested by X-ray photoelectron spectroscopy (XPS) and X-ray powder diffraction analysis (XRPD). Finally, cytotoxicity of the investigated AgNPs has been tested by MTT assay.
Bio-protocol, 2018
Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surg... more Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surgical and pharmacological-based on somatostatin and dopamine agonists-approaches. Although mostly benign tumors, the occurrence of invasive behaviors is often detected resulting in poorer prognosis. The use of primary cultures from human pituitary adenomas represented a significant advancement in the knowledge of the mechanisms of their development and in the definition of the determinants of their pharmacological sensitivity. Moreover, recent studies identified also in pituitary adenomas putative tumor stem cells representing, according to the current hypothesis, the real cellular targets to eradicate most malignancies. In this protocol, we describe the procedure to establish primary cultures from human pituitary adenomas, and how to select, in vitro expand, and phenotypically characterize putative pituitary adenoma stem cells.
Journal of Pharmaceutical and Biomedical Analysis, Sep 1, 2023
Highlights A GC-MS method was developed and validated for the simultaneous determination of hyd... more Highlights A GC-MS method was developed and validated for the simultaneous determination of hydroxyzine and its active metabolite, cetirizine, in whole blood. The developed method provided significant advantages regarding sensitivity and linear dynamic range of related, previously published, methods. The method can be used during the investigation of clinical and forensic toxicology cases. The developed method was successfully applied during the investigation of clinical cases where these antihistamines were detected. The method could also be applied to pharmacokinetic studies concerning these antihistamines. The developed method could be used for the determination of more antihistamines in blood and other biological fluids after proper optimization and validation.
Biomedicines, Feb 16, 2023
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Left Panels: negative controls (stained only with the secondary antibody) for the NFPAs depicted ... more Left Panels: negative controls (stained only with the secondary antibody) for the NFPAs depicted in lanes 1 and 2 of Figure 1 and the GHomas reporeted in lanes 3 and 4. Right Panels: control of specificity for the antibodies in which the staining of the GHoma depicted in the line 3 was abolished preincubating the primary antibodies for SDF1 and CXCR4 with blocking peptides. Supplementary Figure 2 Expression of CXCR4 and SDF1 by IHC in human pituitary GHoma surgical specimen (T = tumor) with adjacent normal pituitary (N= normal).
Frontiers in Endocrinology, Feb 20, 2020
Pituitary adenomas, accounting for 15% of diagnosed intracranial neoplasms, are usually benign an... more Pituitary adenomas, accounting for 15% of diagnosed intracranial neoplasms, are usually benign and pharmacologically and surgically treatable; however, the critical location, mass effects and hormone hypersecretion sustain their significant morbidity. Approximately 35% of pituitary tumors show a less benign course since they are highly proliferative and invasive, poorly resectable, and likely recurring. The latest WHO classification of pituitary tumors includes pituitary transcription factor assessment to determine adenohypophysis cell lineages and accurate designation of adenomas, nevertheless little is known about molecular and cellular pathways which contribute to pituitary tumorigenesis. In malignant tumors the identification of cancer stem cells radically changed the concepts of both tumorigenesis and pharmacological approaches. Cancer stem cells are defined as a subset of undifferentiated transformed cells from which the bulk of cancer cells populating a tumor mass is generated. These cells are able to self-renew, promoting tumor progression and recurrence of malignant tumors, also conferring cytotoxic drug resistance. On the other hand, the existence of stem cells within benign tumors is still debated. The presence of adult stem cells in human and murine pituitaries where they sustain the high plasticity of hormone-producing cells, allowed the hypothesis that putative tumor stem cells might exist in pituitary adenomas, reinforcing the concept that the cancer stem cell model could also be applied to pituitary tumorigenesis. In the last few years, the isolation and phenotypic characterization of putative pituitary adenoma stem-like cells was performed using a wide and heterogeneous variety of experimental models and techniques, although the role of these cells in adenoma initiation and progression is still not completely definite. The assessment of possible pituitary adenoma-initiating cell population would be of extreme relevance to better understand pituitary tumor biology and to identify novel potential diagnostic markers and pharmacological targets. In this review, we summarize the most updated studies focused on the definition of pituitary adenoma stem cell phenotype and functional features, highlighting the biological processes and intracellular pathways potentially involved in driving tumor growth, relapse, and therapy resistance.
Pharmacological Research, Sep 1, 1997
The potential for modulating gene expression by the use of antisense oligonucleotides has become ... more The potential for modulating gene expression by the use of antisense oligonucleotides has become increasingly interesting in recent years. Antisense oligonucleotides are complementary nucleic acid fragments that hybridize to target sequences within RNA to form a DNA-RNA duplex, resulting in the block of translation of messenger RNA into the protein. Advances in chemistry and molecular biology have provided the basis to develop antisense oligodeoxynucleotides and improve their selectivity, stability and specificity of action. The antisense technology has been extensively used in vitro and in vivo as a tool to study the regulatory mechanisms in biologic processes and as potential therapeutic agents in cancer, viral infections and genetic disorders. In the present review, the various approaches for the use of antisense molecules in oncology, virology, genetic and inflammatory diseases are described; several studies, supporting the in vitro and in vivo applications of this technology, are also presented. Moreover, the potential clinical use of antisense therapies is discussed.
bioRxiv (Cold Spring Harbor Laboratory), Dec 31, 2022
Metformin is the first-line drug for type-2 diabetes. Retrospective analyses, based on diabetic p... more Metformin is the first-line drug for type-2 diabetes. Retrospective analyses, based on diabetic patients' clinical data, demonstrate that daily assumption of metformin reduces the incidence of several kinds of solid tumors. Even though it is widely agreed that metformin must be internalized to accomplish its pharmacological activity, direct evidence about metformin membrane permeability and/or the presence of a specific membrane receptor in cancer cells is still missing. Here, we show that the transmembrane form of Chloride Intracellular Channel 1 (tmCLIC1) works as a privileged metformin receptor in glioblastoma stem-like cells. We found that metformin impairs tmCLIC1 activity by a specific binding coordinated by arginine 29. Its mutation, preventing metformin to bind and block tmCLIC1, abolishes the biguanide inhibition of glioblastoma cell proliferation in 2D and 3D models and metformin dependent effect on mitochondrial respiration. In addition, we demonstrate the direct binding between the drug and its target, and by in vivo experiments on zebrafish embryos and mice orthotopically engrafted with glioblastoma cells and treated with metformin, we prove that metformin binding to tmCLIC1 is crucial for metformin antineoplastic effect. Considering tmCLIC1's contribution to glioblastoma progression, the present work provides the fundaments for future development of strategies aimed at improving metformin-tmCLIC1 interaction to further increase metformin therapeutic potential.
Purpose: Hypothalamic or locally produced growth factors and cytokines control pituitary developm... more Purpose: Hypothalamic or locally produced growth factors and cytokines control pituitary development, functioning, and cell division. We evaluated the expression of the chemokine stromal cell–derived factor 1 (SDF1) and its receptor CXCR4 in human pituitary adenomas and normal pituitary tissues and their role in cell proliferation.Experimental Design: The expression of SDF1 and CXCR4 in 65 human pituitary adenomas and 4 human normal pituitaries was determined by reverse transcription-PCR, immunohistochemistry, and confocal immunofluorescence. The proliferative effect of SDF1 was evaluated in eight fibroblast-free human pituitary adenoma cell cultures.Results: CXCR4 mRNA was expressed in 92% of growth hormone (GH)-secreting pituitary adenomas (GHoma) and 81% of nonfunctioning pituitary adenomas (NFPA), whereas SDF1 was identified in 63% and 78% of GHomas and NFPAs, respectively. Immunostaining for CXCR4 and SDF1 showed a strong homogenous labeling in all tumoral cells in both GHomas and NFPAs. In normal tissues, CXCR4 and SDF1 were expressed only in a subset of anterior pituitary cells, with a lower expression of SDF1 compared with its cognate receptor. CXCR4 and SDF1 were not confined to a specific cell population in the anterior pituitary but colocalized with discrete subpopulations of GH-, prolactin-, and adrenocorticorticotropic hormone–secreting cells. Conversely, most of the SDF1-containing cells expressed CXCR4. In six of eight pituitary adenoma primary cultures, SDF1 induced a statistically significant increase in DNA synthesis that was prevented by the treatment with the CXCR4 antagonist AMD3100 or somatostatin.Conclusions: CXCR4 and SDF1 are overexpressed in human pituitary adenomas and CXCR4 activation may contribute to pituitary cell proliferation and, possibly, to adenoma development `in humans.
Journal of Comparative Pathology, 2017
Journal of Clinical Oncology, Jun 1, 2005
Background: Imatinib mesylate (STI-571, Gleevec) a small molecule tyrosine kinase inhibitor direc... more Background: Imatinib mesylate (STI-571, Gleevec) a small molecule tyrosine kinase inhibitor directed against the enzymatic domain of KIT protein, was recently found to produce dramatic clinical responses as monotherapy for metastatic GISTs. However, imatinib resistance can occur determining treatment failure. The present study was performed to analyse the expression of somatostatin receptor (SSTR) subtypes in a series of Gastrointestinal (GI) stromal tumors by immunohistochemistry and possibly identify an additional therapeutic target. Methods: Sixteen cases of GI stromal tumors (8 males and 8 females, age range: 38\u201373) were studied. 11 GISTs, 4 leiomyosarcomas, 1 leiomyoma were included. A gastric leiomyosarcoma was doubly evaluated (primitive tumor and metastatic lung lesion). Immunohistochemical detection of SSTRs was performed on sections (4\u3bcm) of paraffin-embedded tissue stained with polyclonal antibodies directed against the five somatostatin receptor subtypes. Results: We found 7 out of 16 tumors (44%) expressing all SSTRs and 14 out of 16 (87%) tumors positive for at least 3 subtypes. SSTR2A was the most represented subtype in the tumors studied, being expressed in approximately 70% of cases and usually exhibiting an intense labeling. All GISTs apart one resulted positive for more than two SSTR subtypes. SSTR2A was the most strongly expressed subtype in GISTs (72% of cases). All 4 leiomyosarcomas of this series expressed SSTRs, being SSTR4 and SSTR5 positive in all cases. Conclusions: The significant expression of SSTRs showed in this series of GI stromal tumors suggests a potential therapeutic target to be explored alone and/or in combination with other therapeutic agents in the setting of refractory GI stromal tumors
Cells, Dec 13, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Cancer Research
In developed countries, colorectal cancer is the third leading cause of mortality. Despite signif... more In developed countries, colorectal cancer is the third leading cause of mortality. Despite significant advances in therapeutic approaches, metastases remain the primary concern in the treatment of colorectal cancer. Ion channels have an important role in the development of aggressive behavior, but the majority of them are involved in regular physiological activities. Chloride Intracellular Channel 1 (CLIC1) is a promising contender among the new possible biomarkers. CLIC1 is expressed as a cytosolic monomer by all cell types during physiological homeostasis. The main characteristic of this protein is that it can exist in both cytosolic and transmembrane forms (tmCLIC1), with the latter being involved in a variety of clinical conditions, including cancer. CLIC1 mRNA overexpression in colorectal cancer cells has been linked to a poor prognosis. The primary goal of this study is to understand the role of tmCLIC1 in the carcinogenesis, invasion, and migratory potential of colorectal can...
Neuro-Oncology
Background Meningiomas are mainly benign brain tumors, although about 20% of histologically benig... more Background Meningiomas are mainly benign brain tumors, although about 20% of histologically benign cases are clinically aggressive and recur after resection. We hypothesize that meningioma brain invasiveness and recurrence may be related to the presence of cancer stem cells and their high responsiveness to the CXCL12-CXCR4/CXCR7 chemokine axis. The aim of this study was to isolate meningioma stem cells from human samples, characterize them for biological features related to malignant behavior, and to identify the role of CXCR4/CXCR7 in these processes. Methods Meningioma stem cells were isolated from patient-derived primary cultures in stem cell-permissive conditions, and characterized for phenotype, self-renewal, proliferation and migration rates, vasculogenic mimicry (VM), and in vivo tumorigenesis, in comparison with differentiated meningioma cells and stem-like cells isolated from normal meninges. These cell populations were challenged with CXCL12 and CXCL11 and receptor antagon...
Effect of IAA on apoptosis in GB CSCs.
Current density/voltage relationships of CLIC1 current in CSCs
Glioblastoma (GB) is the most lethal, aggressive, and diffuse brain tumor. The main challenge for... more Glioblastoma (GB) is the most lethal, aggressive, and diffuse brain tumor. The main challenge for successful treatment is targeting the cancer stem cell (CSC) subpopulation responsible for tumor origen, progression, and recurrence. Chloride Intracellular Channel 1 (CLIC1), highly expressed in CSCs, is constitutively present in the plasma membrane where it is associated with chloride ion permeability. In vitro, CLIC1 inhibition leads to a significant arrest of GB CSCs in G1 phase of the cell cycle. Furthermore, CLIC1 knockdown impairs tumor growth in vivo. Here, we demonstrate that CLIC1 membrane localization and function is specific for GB CSCs. Mesenchymal stem cells (MSC) do not show CLIC1-associated chloride permeability, and inhibition of CLIC1 protein function has no influence on MSC cell-cycle progression. Investigation of the basic functions of GB CSCs reveals a constitutive state of oxidative stress and cytoplasmic alkalinization compared with MSCs. Both intracellular oxidat...
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Papers by Federica Barbieri