Oncotarget

Research Papers:

Identification of BAG3 target proteins in anaplastic thyroid cancer cells by proteomic analysis

Francesca Galdiero _, Anna Maria Bello, Anna Spina, Anna Capiluongo, Sophie Liuu, Margot De Marco, Alessandra Rosati, Mario Capunzo, Maria Napolitano, Emilia Vuttariello, Mario Monaco, Daniela Califano, Maria Caterina Turco, Gennaro Chiappetta, Joëlle Vinh and Giovanni Chiappetta

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Oncotarget. 2018; 9:8016-8026. https://doi.org/10.18632/oncotarget.23858

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Abstract

Francesca Galdiero1, Anna Maria Bello1, Anna Spina1, Anna Capiluongo1, Sophie Liuu2, Margot De Marco3, Alessandra Rosati3,4, Mario Capunzo4, Maria Napolitano1, Emilia Vuttariello1, Mario Monaco1, Daniela Califano1, Maria Caterina Turco3,4,5, Gennaro Chiappetta1, Joëlle Vinh2 and Giovanni Chiappetta2

1Functional Genomic Unit, Istituto Nazionale Tumori–IRCCS–Fondazione G.Pascale, Napoli, Italia

2ESPCI ParisTech, Spectrométrie de Masse Biologique et Protéomique (SMBP), USR3149 CNRS, Paris, France

3Biouniversa s.r.l., University of Salerno, Fisciano, Italy

4Department of Medicine, Surgery and Dentistry "Scuola Medica Salernitana", University of Salerno, Baronissi (SA), Italy

5"SS. Giovanni di Dio e Ruggi d'Aragona-Schola Medica Salernitana", University of Salerno Hospital, Salerno, Italy

Correspondence to:

Gennaro Chiappetta, email: g.chiappetta@istitutotumori.na.it

Keywords: BAG3; anaplastic thyroid cancer; SILAC; CAV1; SERPINB2

Received: May 18, 2017     Accepted: October 30, 2017     Published: January 03, 2018

ABSTRACT

BAG3 protein is an apoptosis inhibitor and is highly expressed in Anaplastic Thyroid Cancer. We investigated the entire set of proteins modulated by BAG3 silencing in the human anaplastic thyroid 8505C cancer cells by using the Stable-Isotope Labeling by Amino acids in Cell culture strategy combined with mass spectrometry analysis. By this approach we identified 37 up-regulated and 54 down-regulated proteins in BAG3-silenced cells. Many of these proteins are reportedly involved in tumor progression, invasiveness and resistance to therapies. We focused our attention on an oncogenic protein, CAV1, and a tumor suppressor protein, SERPINB2, that had not previously been reported to be modulated by BAG3. Their expression levels in BAG3-silenced cells were confirmed by qRT-PCR and western blot analyses, disclosing two novel targets of BAG3 pro-tumor activity. We also examined the dataset of proteins obtained by the quantitative proteomics analysis using two tools, Downstream Effect Analysis and Upstream Regulator Analysis of the Ingenuity Pathways Analysis software. Our analyses confirm the association of the proteome profile observed in BAG3-silenced cells with an increase in cell survival and a decrease in cell proliferation and invasion, and highlight the possible involvement of four tumor suppressor miRNAs and TP53/63 proteins in BAG3 activity.


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