title: Anti-Neuroblastoma Properties of a Recombinant Sunflower Lectin.
abstract: According to their sugar recognition specificity, plant lectins are proposed as bioactive proteins with potential in cancer treatment and diagnosis. Helja is a mannose-specific jacalin-like lectin from sunflower which was shown to inhibit the growth of certain fungi. Here, we report its recombinant expression in a prokaryotic system and its activity in neurobalstoma cells. Helja coding sequence was fused to the pET-32 EK/LIC, the enterokinase/Ligation-independent cloning vector and a 35 kDa protein was obtained inEscherichia colirepresenting Helja coupled to thioredoxin (Trx). The identity of this protein was verified using anti-Helja antibodies. This chimera, named Trx-rHelja, was enriched in the soluble bacterial extracts and was purified using Ni+2-Sepharose and d-mannose-agarose chromatography. Trx-rHelja and the enterokinase-released recombinant Helja (rHelja) both displayed toxicity on human SH-SY5Y neuroblastomas. rHelja decreased the viability of these tumor cells by 75% according to the tetrazolium reduction assay, and microscopic analyses revealed that the cell morphology was disturbed. Thus, the stellate cells of the monolayer became spheroids and were isolated. Our results indicate that rHelja is a promising tool for the development of diagnostic or therapeutic methods for neuroblastoma cells, the most common solid tumors in childhood.
authors.affiliation: Instituto de Investigaciones Biológicas (IIB), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, Funes 3250, 7600 Mar del Plata, Argentina. mpinedo@mdp.edu.ar.
authors.forename: Marcela
authors.initials: M
authors.lastname: Pinedo
authors.affiliation: Instituto de Investigaciones Biológicas (IIB), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, Funes 3250, 7600 Mar del Plata, Argentina. mgenoula@gmail.com.
authors.forename: Melanie
authors.initials: M
authors.lastname: Genoula
authors.affiliation: Instituto de Investigaciones Biológicas (IIB), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, Funes 3250, 7600 Mar del Plata, Argentina. mxsilveyra@gmail.com.
authors.forename: María Ximena
authors.initials: MX
authors.lastname: Silveyra
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes 28013-602, Brazil. andre@uenf.br.
authors.forename: André
authors.initials: A
authors.lastname: De Oliveira Carvalho
authors.affiliation: Instituto de Investigaciones Biológicas (IIB), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, Funes 3250, 7600 Mar del Plata, Argentina. mregente@mdp.edu.ar.
authors.forename: Mariana
authors.initials: M
authors.lastname: Regente
authors.affiliation: Instituto de Investigaciones Biológicas (IIB), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, Funes 3250, 7600 Mar del Plata, Argentina. maris03_5@hotmail.com.
authors.forename: Marianela
authors.initials: M
authors.lastname: Del Río
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes 28013-602, Brazil. julliah@hotmail.com.
authors.forename: Júlia
authors.initials: J
authors.lastname: Ribeiro Soares
authors.affiliation: Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes 28013-602, Brazil. valmg@uenf.br.
authors.forename: Valdirene
authors.initials: V
authors.lastname: Moreira Gomes
authors.affiliation: Instituto de Investigaciones Biológicas (IIB), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, Funes 3250, 7600 Mar del Plata, Argentina. ldelacan@mdp.edu.ar.
authors.forename: Laura
authors.initials: L
authors.lastname: De La Canal
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006368:Helianthus / Q000737:chemistry*; D006801:Humans; D009447:Neuroblastoma / Q000188:drug therapy / Q000473:pathology; D037121:Plant Lectins / Q000302:isolation & purification / Q000494:pharmacology*; D011994:Recombinant Proteins
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D037121:Plant Lectins; D011994:Recombinant Proteins; C043696:jacalin
keywords: Helianthus annuus; Helja; agglutinin; jacalin; neuroblastoma; recombinant lectin; tumor toxicity