9 results on '"Alessandra di Gennaro"'
Search Results
2. A p53/miR-30a/ZEB2 axis controls triple negative breast cancer aggressiveness
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Tiziana Perin, Antonella Zucchetto, Alessandra di Gennaro, Herman P. Spaink, Michela Armellin, Valentina Damiano, Manuela Santarosa, Roberta Maestro, B. Ewa Snaar-Jagalska, Claudio Doglioni, Michela Guardascione, Giulia Brisotto, Di Gennaro, A., Damiano, V., Brisotto, G., Armellin, M., Perin, T., Zucchetto, A., Guardascione, M., Spaink, H. P., Doglioni, C., Snaar-Jagalska, B. E., Santarosa, M., and Maestro, R.
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0301 basic medicine ,In silico ,Triple Negative Breast Neoplasms ,Drug resistance ,Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Transcription (biology) ,Cell Line, Tumor ,microRNA ,medicine ,Humans ,Epigenetics ,Molecular Biology ,Lymph node ,Transcription factor ,Triple-negative breast cancer ,Zinc Finger E-box Binding Homeobox 2 ,Correction ,Cell Biology ,Middle Aged ,MicroRNAs ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Tumor Suppressor Protein p53 - Abstract
Inactivation of p53 contributes significantly to the dismal prognosis of breast tumors, most notably triple-negative breast cancers (TNBCs). How the relief from p53 tumor suppressive functions results in tumor cell aggressive behavior is only partially elucidated. In an attempt to shed light on the implication of microRNAs in this context, we discovered a new signaling axis involving p53, miR-30a and ZEB2. By an in silico approach we identified miR-30a as a putative p53 target and observed that in breast tumors reduced miR-30a expression correlated with p53 inactivation, lymph node positivity and poor prognosis. We demonstrate that p53 binds the MIR30A promoter and induces the transcription of both miRNA strands 5p and 3p. Both miR-30a-5p and -3p showed the capacity of targeting ZEB2, a transcription factor involved in epithelial-mesenchymal transition (EMT), tumor cell migration and drug resistance. Intriguingly, we found that p53 does restrain ZEB2 expression via miR-30a. Finally, we provide evidence that the new p53/miR-30a/ZEB2 axis controls tumor cell invasion and distal spreading and impinges upon miR-200c expression. Overall, this study highlights the existence of a novel axis linking p53 to EMT via miR-30a, and adds support to the notion that miRNAs represent key elements of the complex network whereby p53 inactivation affects TNBC clinical behavior.
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- 2018
3. Epigenetic silencing of miR-200c in breast cancer is associated with aggressiveness and is modulated by ZEB1
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Alessandra di Gennaro, Silvia Borgna, Manuela Santarosa, Roberta Maestro, Valentina Damiano, Michela Guardascione, Giulia Brisotto, Tiziana Perin, and Michela Armellin
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0301 basic medicine ,Cancer Research ,Methylation ,Biology ,Chromatin remodeling ,Demethylating agent ,Chromatin ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,Histone ,chemistry ,030220 oncology & carcinogenesis ,microRNA ,DNA methylation ,Genetics ,Cancer research ,biology.protein ,Gene silencing - Abstract
Loss of expression of miR-200 family members has been implicated in cellular plasticity, a phenomenon that accounts for epithelial-to-mesenchymal transition (EMT) and stem-like features of many carcinomas and is considered a major cause of tumor aggressiveness and drug resistance. Nevertheless, the mechanisms of miR-200 downregulation in breast cancer are still largely unknown. Here we show that miR-200c expression inversely correlates with miR-200c/miR-141 locus methylation in triple-negative breast tumors (TNBC). Importantly, low levels of miR-200c expression and high levels of miR-200c/miR-141 locus methylation associated with lymph node metastasis. Moreover, miR-200c/miR-141 locus methylation was significantly related to high expression of ZEB1 in two independent TNBC series. Silencing of ZEB1 in vitro reduced miR-200c/miR-141 DNA methylation and, concurrently, decreased histone H3K9 trimethylation. This chromatin modifications were paralleled by an increase in the expression of both miR-200c and E-cadherin. Similar effects were achieved by treatment with a demethylating agent. Our data suggest that gene methylation is an important element in the regulation of the miR-200c/ZEB1 axis and that chromatin remodeling of the miR-200c/miR-141 locus is affected by ZEB1 and, thus, contributes to ZEB1-induced cellular plasticity. © 2016 Wiley Periodicals, Inc.
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- 2016
4. Correction to: A p53/miR-30a/ZEB2 axis controls triple negative breast cancer aggressiveness
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Tiziana Perin, Roberta Maestro, Claudio Doglioni, Herman P. Spaink, Michela Guardascione, Alessandra di Gennaro, Michela Armellin, Valentina Damiano, Manuela Santarosa, Giulia Brisotto, Antonella Zucchetto, and B. Ewa Snaar-Jagalska
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0301 basic medicine ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,business.industry ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Medicine ,Cell Biology ,business ,Molecular Biology ,Triple-negative breast cancer - Abstract
Since publication of the article, the authors were notified by ATCC that the cell line HCC1395 (ATCC® CRL-2324™ Lot 62235652) suffered a “low level of cell line cross-contamination” with another cell line.
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- 2019
5. Functional Analysis of the MurinePax8Promoter Reveals Autoregulation and the Presence of a Novel Thyroid-Specific DNA-Binding Activity
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Olga Spadaro, Mario De Felice, Maria Giuseppina Baratta, Alessandra di Gennaro, Roberto Di Lauro, di Gennaro, A, Spadaro, O, Baratta, Mg, DE FELICE, Mario, and DI LAURO, Roberto
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Transcriptional Activation ,endocrine system ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Thyroid Gland ,Mutagenesis (molecular biology technique) ,Biology ,Mice ,PAX8 Transcription Factor ,Endocrinology ,Gene expression ,medicine ,Animals ,Homeostasis ,Paired Box Transcription Factors ,Promoter Regions, Genetic ,Gene ,Binding Sites ,Thyroid ,Promoter ,Transfection ,Molecular biology ,medicine.anatomical_structure ,Mutagenesis ,PAX8 ,Haploinsufficiency - Abstract
Background: Organogenesis of the thyroid gland requires the Pax8 protein. Absence or reduction of Pax8 results in congenital hypothyroidism in animal models and humans, respectively. This study aims at elucidating the regulatory mechanism leading to the expression of Pax8 in thyroid cells. Methods: The murine Pax8 gene promoter was functionally dissected by mutagenesis and transfection in the thyroid cell line FRTL-5. Nuclear factors important for thyroid-specific gene expression were identified by DNA-binding assays. Results: We show that Pax8 binds to and controls the expression of its own promoter. Furthermore, we identify a novel, thyroid-specific, DNA-binding activity (denominated nTTF [for novel Thyroid Transcription Factor]) that recognizes a specific region of the Pax8 promoter. Conclusions: The Pax8 promoter appears to be autoregulated, a feature that might be responsible for the haploinsufficiency displayed by this gene. © Copyright 2013, Mary Ann Liebert, Inc. 2013.
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- 2013
6. Epigenetic silencing of miR-200c in breast cancer is associated with aggressiveness and is modulated by ZEB1
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Valentina, Damiano, Giulia, Brisotto, Silvia, Borgna, Alessandra, di Gennaro, Michela, Armellin, Tiziana, Perin, Michela, Guardascione, Roberta, Maestro, and Manuela, Santarosa
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Epithelial-Mesenchymal Transition ,Blotting, Western ,Apoptosis ,Triple Negative Breast Neoplasms ,Real-Time Polymerase Chain Reaction ,Epigenesis, Genetic ,Immunoenzyme Techniques ,Cell Movement ,Biomarkers, Tumor ,Tumor Cells, Cultured ,Humans ,Neoplasm Invasiveness ,Gene Silencing ,RNA, Messenger ,Cell Proliferation ,Neoplasm Staging ,Retrospective Studies ,Reverse Transcriptase Polymerase Chain Reaction ,Zinc Finger E-box-Binding Homeobox 1 ,DNA Methylation ,Middle Aged ,Prognosis ,Combined Modality Therapy ,Gene Expression Regulation, Neoplastic ,Survival Rate ,MicroRNAs ,Lymphatic Metastasis ,Female ,Neoplasm Grading ,Follow-Up Studies - Abstract
Loss of expression of miR-200 family members has been implicated in cellular plasticity, a phenomenon that accounts for epithelial-to-mesenchymal transition (EMT) and stem-like features of many carcinomas and is considered a major cause of tumor aggressiveness and drug resistance. Nevertheless, the mechanisms of miR-200 downregulation in breast cancer are still largely unknown. Here we show that miR-200c expression inversely correlates with miR-200c/miR-141 locus methylation in triple-negative breast tumors (TNBC). Importantly, low levels of miR-200c expression and high levels of miR-200c/miR-141 locus methylation associated with lymph node metastasis. Moreover, miR-200c/miR-141 locus methylation was significantly related to high expression of ZEB1 in two independent TNBC series. Silencing of ZEB1 in vitro reduced miR-200c/miR-141 DNA methylation and, concurrently, decreased histone H3K9 trimethylation. This chromatin modifications were paralleled by an increase in the expression of both miR-200c and E-cadherin. Similar effects were achieved by treatment with a demethylating agent. Our data suggest that gene methylation is an important element in the regulation of the miR-200c/ZEB1 axis and that chromatin remodeling of the miR-200c/miR-141 locus is affected by ZEB1 and, thus, contributes to ZEB1-induced cellular plasticity. © 2016 Wiley Periodicals, Inc.
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- 2016
7. Abstract 500: A p53/miR-30a/ZEB2 axis controls basal-like/triple-negative breast cancer aggressiveness
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Manuela Santarosa, Giulia Brisotto, Claudio Doglioni, Michela Guardascione, Alessandra di Gennaro, Herman P. Spaink, Tiziana Perin, Michela Armellin, Valentina Damiano, Antonella Zucchetto, Ewa Snaar-Jagalska, and Roberta Maestro
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Cancer Research ,Basal (phylogenetics) ,Oncology ,business.industry ,Cancer research ,Medicine ,business ,Triple-negative breast cancer - Abstract
Inactivation of p53 plays a major role in the poor prognosis of basal-like /triple-negative breast cancers (TNBC). However, how p53 inactivation impinges upon TNBC aggressive phenotype is only partially defined. Here we report the identification of a novel axis involving p53, miR30a and ZEB2. We provide evidence that p53 affects the expression of ZEB2, a transcription factor involved in epithelial-mesenchymal transition (EMT), by relying on miR-30a. In particular, we found that p53 regulates miR-30a expression by direct promoter binding and that miR30a targets ZEB2. Finally, we provide evidence that the new p53/miR-30a/ZEB2 axis controls tumor cell invasion and migration and that reduced miR-30a expression correlates with p53 inactivation and dismal prognosis in human breast cancers. Overall, this study highlights the existence of a novel axis linking p53 to EMT via miR-30a, and adds a novel element to the complex network whereby p53 inactivation impinge upon tumor aggressiveness. Citation Format: Valentina Damiano, Alessandra di Gennaro, Giulia Brisotto, Michela Armellin, Tiziana Perin, Antonella Zucchetto, Michela Guardascione, Herman P. Spaink, Claudio Doglioni, Ewa Snaar-Jagalska, Manuela Santarosa, Roberta Maestro. A p53/miR-30a/ZEB2 axis controls basal-like/triple-negative breast cancer aggressiveness [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 500.
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- 2018
8. Comparative genomics reveals a functional thyroid-specific element in the far upstream region of the PAX8 gene
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Tiziana de Cristofaro, Mariastella Zannini, Roberto Nitsch, Roberto Di Lauro, Alessandra di Gennaro, Mario De Felice, Serena Abbondante, Valeria Di Dato, Nitsch, R., Di Dato, V., di Gennaro, A., de Cristofaro, T., Abbondante, S., De Felice, M., Zannini, M., and Di Lauro, R.
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Transcription, Genetic ,endocrine system diseases ,PROMOTER ,5' Flanking Region ,medicine.medical_treatment ,Thyroid Gland ,Mice ,0302 clinical medicine ,Paired Box Transcription Factors ,TRANSCRIPTION FACTOR ,Conserved Sequence ,Phylogeny ,Genetics ,Regulation of gene expression ,0303 health sciences ,biology ,HOMEODOMAIN ,Genomics ,DNA-Binding Proteins ,DIFFERENTIATION ,Enhancer Elements, Genetic ,Organ Specificity ,030220 oncology & carcinogenesis ,RECEPTOR ACTIVATOR ,Research Article ,Biotechnology ,endocrine system ,lcsh:QH426-470 ,lcsh:Biotechnology ,5' flanking region ,BAC DNA ,ENHANCER ,PAX8 Transcription Factor ,03 medical and health sciences ,Thyroid peroxidase ,lcsh:TP248.13-248.65 ,Sequence Homology, Nucleic Acid ,medicine ,Animals ,Humans ,Enhancer ,Transcription factor ,030304 developmental biology ,RECOGNITION ,lcsh:Genetics ,Gene Expression Regulation ,CELLS ,biology.protein ,Thyroglobulin ,PAX8 ,Sequence Alignment ,TISSUE-SPECIFIC EXPRESSION ,Transcription Factors ,FOXE1 - Abstract
Background The molecular mechanisms leading to a fully differentiated thyrocite are still object of intense study even if it is well known that thyroglobulin, thyroperoxidase, NIS and TSHr are the marker genes of thyroid differentiation. It is also well known that Pax8, TTF-1, Foxe1 and Hhex are the thyroid-enriched transcription factors responsible for the expression of the above genes, thus are responsible for the differentiated thyroid phenotype. In particular, the role of Pax8 in the fully developed thyroid gland was studied in depth and it was established that it plays a key role in thyroid development and differentiation. However, to date the bases for the thyroid-enriched expression of this transcription factor have not been unraveled yet. Here, we report the identification and characterization of a functional thyroid-specific enhancer element located far upstream of the Pax8 gene. Results We hypothesized that regulatory cis-acting elements are conserved among mammalian genes. Comparison of a genomic region extending for about 100 kb at the 5'-flanking region of the mouse and human Pax8 gene revealed several conserved regions that were tested for enhancer activity in thyroid and non-thyroid cells. Using this approach we identified one putative thyroid-specific regulatory element located 84.6 kb upstream of the Pax8 transcription start site. The in silico data were verified by promoter-reporter assays in thyroid and non-thyroid cells. Interestingly, the identified far upstream element manifested a very high transcriptional activity in the thyroid cell line PC Cl3, but showed no activity in HeLa cells. In addition, the data here reported indicate that the thyroid-enriched transcription factor TTF-1 is able to bind in vitro and in vivo the Pax8 far upstream element, and is capable to activate transcription from it. Conclusions Results of this study reveal the presence of a thyroid-specific regulatory element in the 5' upstream region of the Pax8 gene. The identification of this regulatory element represents the first step in the investigation of upstream regulatory mechanisms that control Pax8 transcription during thyroid differentiation and are relevant to further studies on Pax8 as a candidate gene for thyroid dysgenesis.
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- 2010
9. An improved sequencing-based strategy to estimate locus-specific DNA methylation
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Roberta Maestro, Alessandra di Gennaro, Tiziana Perin, Giulia Brisotto, Manuela Santarosa, Michela Armellin, and Valentina Damiano
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Cancer Research ,Method ,Locus (genetics) ,Breast Neoplasms ,Biology ,Bisulfite treatment ,chemistry.chemical_compound ,Cell Line, Tumor ,microRNA ,Genetics ,Sequencing ,Humans ,Promoter Regions, Genetic ,Gene ,Cancer ,CDH1 ,E-cadherin ,Promoter ,Methylation ,Sequence Analysis, DNA ,DNA Methylation ,MicroRNAs ,chemistry ,CpG site ,Technical Advance ,Oncology ,Genetic Loci ,DNA methylation ,Illumina Methylation Assay ,Female ,DNA-methylation ,DNA ,miR-200c/miR-141 locus - Abstract
Background DNA methylation is an important epigenetic mechanism of transcriptional control that plays an essential role in several cellular functions. Aberrant DNA methylation in cancer has been frequently associated with downregulation of microRNAs and protein coding genes, such as miR-200c/miR-141 cluster and E-cadherin. Current strategies to assess DNA methylation, including bisulfite treatment-based assays, tend to be time-consuming and may be quite expensive when a precise appraisal is required. The Sanger-sequencing of the amplified bisulfite-treated DNA (BSP) might represent a practical option to measure DNA methylation at single CpG resolution. However, this strategy often produces noisy data, which affects accurate quantification. Here we propose an improved, reliable and cost-effective BSP-based protocol that allows proper DNA methylation assessment. Methods Our strategy, named normalized-BSP (NBSP), takes advantage of tailed C-balanced primers and a normalization procedure based on C/T ratio to overcome BSP-associated noise problems and nucleotide signal unbalance. NBSP was applied to estimate miR-200c/miR-141 locus methylation in serial dilution experiments and was compared to conventional methods. Besides, it was applied in the analysis of FFPE breast cancer samples and further validated in the context of the E-cadherin promoter. Results NBSP strategy outperformed conventional BSP in the estimate of the fraction of methylated cytosine in serial dilution experiments, providing data in agreement with the widely used but cumbersome cloning-based protocol. This held true for both miR-200c/miR-141 locus and E-cadherin promoter analyses. Moreover, the miR-200c/miR-141 locus methylation reflected the decrease in miRNA expression both in breast cancer cell lines and in the FFPE samples. Conclusions NBSP is a rapid and economical method to estimate the extent of methylation at each CpG of a given locus. Notably, NBSP works efficiently on FFPE samples, thus disclosing the perspective of its application also in the diagnostic setting. Electronic supplementary material The online version of this article (doi:10.1186/s12885-015-1646-6) contains supplementary material, which is available to authorized users.
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