Oncotarget

Research Papers:

Construction and characterization of a new TRAIL soluble form, active at picomolar concentrations

Matias Eliseo Melendez, Renato José Silva-Oliveira, Anna Luiza Silva Almeida Vicente, Lidia Maria Rebolho Batista Arantes, Ana Carolina de Carvalho, Alberto Luis Epstein, Rui Manuel Reis and André Lopes Carvalho _

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Oncotarget. 2018; 9:27233-27241. https://doi.org/10.18632/oncotarget.25519

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Abstract

Matias Eliseo Melendez1, Renato José Silva-Oliveira1, Anna Luiza Silva Almeida Vicente1, Lidia Maria Rebolho Batista Arantes1, Ana Carolina de Carvalho1, Alberto Luis Epstein2, Rui Manuel Reis1,3,4 and André Lopes Carvalho1

1Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos, São Paulo, Brazil

2UMR1179, INSERM-UVSQ, Handicap Neuromusculaire, Biotherapie et Pharmacologie Appliquées, Université de Versailles-Saint Quentin en Yvelines, Versailles, France

3Life and Health Sciences Research Institute (ICVS), Health Sciences School, University of Minho, Braga, Portugal

4ICVS/3B’s-PT Government Associate Laboratory, Braga/Guimarães, Portugal

Correspondence to:

André Lopes Carvalho, email: carvalhoal@gmail.com

Keywords: TRAIL; apoptosis; cancer treatment; amplicon vectors

Received: August 12, 2016    Accepted: May 14, 2018    Published: June 05, 2018

ABSTRACT

Apoptosis induction has emerged as a treatment option for anticancer therapy. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a type II transmembrane protein, is a potent and specific pro-apoptotic protein ligand, which activates the extrinsic apoptosis pathway of the cell death receptors. Here we describe the construction and characterization of a new soluble TRAIL, sfTRAIL, stabilized with the trimerization Foldon domain from the Fibritin protein of the bacteriophage T4. Supernatants of 0.22 μM-filtered supernatants were produced in Vero-transduced cells with HSV1-derived viral amplicon vectors. Experiments were undertaken in two known TRAIL-sensitive (U373 and MDA.MB.231) and two TRAIL-resistant (MCF7 and A549) cell lines, to determine (i) whether the sfTRAIL protein is synthetized and, (ii) whether sfTRAIL could induce receptor-mediated apoptosis. Our results showed that sfTRAIL was able to induce apoptosis at concentrations as low as 1899.29 pg/mL (27.71 pM), independently of caspase-9 activation, and reduction in cell viability at 998.73 fM.


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