Cargando…

Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma

Since the current treatments have not resulted in the desired outcomes for melanoma patients, there is a need to identify more effective medications. Together with other snake venom proteins, cytotoxin-II has shown promising results in tumoral cells. In this study, recombinant cytotoxin-II (rCTII) w...

Descripción completa

Detalles Bibliográficos
Autores principales: Derakhshani, Afshin, Silvestris, Nicola, Hemmat, Nima, Asadzadeh, Zahra, Abdoli Shadbad, Mahdi, Nourbakhsh, Niloufar Sadat, Mobasheri, Leila, Vahedi, Parviz, Shahmirzaie, Morteza, Brunetti, Oronzo, Safarpour, Hossein, Baradaran, Behzad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7663949/
https://www.ncbi.nlm.nih.gov/pubmed/33167431
http://dx.doi.org/10.3390/molecules25215148
_version_ 1783609744960782336
author Derakhshani, Afshin
Silvestris, Nicola
Hemmat, Nima
Asadzadeh, Zahra
Abdoli Shadbad, Mahdi
Nourbakhsh, Niloufar Sadat
Mobasheri, Leila
Vahedi, Parviz
Shahmirzaie, Morteza
Brunetti, Oronzo
Safarpour, Hossein
Baradaran, Behzad
author_facet Derakhshani, Afshin
Silvestris, Nicola
Hemmat, Nima
Asadzadeh, Zahra
Abdoli Shadbad, Mahdi
Nourbakhsh, Niloufar Sadat
Mobasheri, Leila
Vahedi, Parviz
Shahmirzaie, Morteza
Brunetti, Oronzo
Safarpour, Hossein
Baradaran, Behzad
author_sort Derakhshani, Afshin
collection PubMed
description Since the current treatments have not resulted in the desired outcomes for melanoma patients, there is a need to identify more effective medications. Together with other snake venom proteins, cytotoxin-II has shown promising results in tumoral cells. In this study, recombinant cytotoxin-II (rCTII) was expressed in SHuffle(®) T7 Express cells, while the epitope mapping of rCTII was performed to reveal the antibody-binding regions of rCTII. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to assess the viability of SK-MEL-3 and HFF-2 cells after treating these cells with rCTII. The qRT-PCR was performed to evaluate the expression levels of matrix metallopeptidase 3 (MMP-3), SMAD2, SMAD3, caspase-8, caspase-9, and miR-214 in order to reveal the rCTII-induced signaling pathways in melanoma. Our results have shown that two regions of amino acids, 6–16 and 19–44, as predicted epitopes of this toxin, are essential for understanding the toxicity of rCTII. Treating the melanoma cells with rCTII substantially inhibited the transforming growth factor-beta (TGF-β)–SMAD signaling pathway and down-regulated the expression of MMP-3 and miR-214 as well. This cytotoxin also restored apoptosis mainly via the intrinsic pathway. The down-regulation of MMP-3 and miR-214 might be associated with the anti-metastatic property of rCTII in melanoma. The inhibitory effect of rCTII on the TGF-β signaling pathway might be associated with increased apoptosis and decreased cancer cell proliferation. It is interesting to see that the IC50 value of rCTII has been lower in the melanoma cells than non-tumoral cells, which may indicate its potential effects as a drug. In conclusion, rCTII, as a novel medication, might serve as a potent and efficient anticancer drug in melanoma.
format Online
Article
Text
id pubmed-7663949
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-76639492020-11-14 Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma Derakhshani, Afshin Silvestris, Nicola Hemmat, Nima Asadzadeh, Zahra Abdoli Shadbad, Mahdi Nourbakhsh, Niloufar Sadat Mobasheri, Leila Vahedi, Parviz Shahmirzaie, Morteza Brunetti, Oronzo Safarpour, Hossein Baradaran, Behzad Molecules Article Since the current treatments have not resulted in the desired outcomes for melanoma patients, there is a need to identify more effective medications. Together with other snake venom proteins, cytotoxin-II has shown promising results in tumoral cells. In this study, recombinant cytotoxin-II (rCTII) was expressed in SHuffle(®) T7 Express cells, while the epitope mapping of rCTII was performed to reveal the antibody-binding regions of rCTII. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to assess the viability of SK-MEL-3 and HFF-2 cells after treating these cells with rCTII. The qRT-PCR was performed to evaluate the expression levels of matrix metallopeptidase 3 (MMP-3), SMAD2, SMAD3, caspase-8, caspase-9, and miR-214 in order to reveal the rCTII-induced signaling pathways in melanoma. Our results have shown that two regions of amino acids, 6–16 and 19–44, as predicted epitopes of this toxin, are essential for understanding the toxicity of rCTII. Treating the melanoma cells with rCTII substantially inhibited the transforming growth factor-beta (TGF-β)–SMAD signaling pathway and down-regulated the expression of MMP-3 and miR-214 as well. This cytotoxin also restored apoptosis mainly via the intrinsic pathway. The down-regulation of MMP-3 and miR-214 might be associated with the anti-metastatic property of rCTII in melanoma. The inhibitory effect of rCTII on the TGF-β signaling pathway might be associated with increased apoptosis and decreased cancer cell proliferation. It is interesting to see that the IC50 value of rCTII has been lower in the melanoma cells than non-tumoral cells, which may indicate its potential effects as a drug. In conclusion, rCTII, as a novel medication, might serve as a potent and efficient anticancer drug in melanoma. MDPI 2020-11-05 /pmc/articles/PMC7663949/ /pubmed/33167431 http://dx.doi.org/10.3390/molecules25215148 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Derakhshani, Afshin
Silvestris, Nicola
Hemmat, Nima
Asadzadeh, Zahra
Abdoli Shadbad, Mahdi
Nourbakhsh, Niloufar Sadat
Mobasheri, Leila
Vahedi, Parviz
Shahmirzaie, Morteza
Brunetti, Oronzo
Safarpour, Hossein
Baradaran, Behzad
Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma
title Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma
title_full Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma
title_fullStr Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma
title_full_unstemmed Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma
title_short Targeting TGF-β-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma
title_sort targeting tgf-β-mediated smad signaling pathway via novel recombinant cytotoxin ii: a potent protein from naja naja oxiana venom in melanoma
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7663949/
https://www.ncbi.nlm.nih.gov/pubmed/33167431
http://dx.doi.org/10.3390/molecules25215148
work_keys_str_mv AT derakhshaniafshin targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT silvestrisnicola targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT hemmatnima targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT asadzadehzahra targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT abdolishadbadmahdi targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT nourbakhshniloufarsadat targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT mobasherileila targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT vahediparviz targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT shahmirzaiemorteza targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT brunettioronzo targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT safarpourhossein targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma
AT baradaranbehzad targetingtgfbmediatedsmadsignalingpathwayvianovelrecombinantcytotoxiniiapotentproteinfromnajanajaoxianavenominmelanoma