Purpose: Tumor cells generally exhibit higher levels of reactive oxygen species (ROS), however, when stressed, tumor cells can undergo a process of ‘Redox Resetting’ to acquire a new redox balance with stronger antioxidant systems that enable cancer cells to become resistant to radiation therapy (RT). Here, we describe how RT affects the oxidant/antioxidant balance in human embryonal (RD) and alveolar (RH30) rhabdomyosarcoma (RMS) cell lines, investigating on the molecular mechanisms involved. Methods: Radiations were delivered using an x-6 MV photon linear accelerator and their effects were assessed by vitality and clonogenic assays. The expression of specific antioxidant-enzymes, such as Superoxide Dismutases (SODs), Catalase (CAT) and Glutathione Peroxidases 4 (GPx4), miRNAs (miR-22, -126, -210, -375, -146a, -34a) and the transcription factor NRF2 was analyzed by quantitative polymerase chain reaction (q-PCR) and western blotting. RNA interference experiments were performed to evaluate the role of NRF2. Results: Doses of RT higher than 2 Gy significantly affected RMS clonogenic ability by increasing ROS production. RMS rapidly and efficiently brought back ROS levels by up-regulating the gene expression of antioxidant enzymes, miRNAs as well as of NRF2. Silencing of NRF2 restrained the RMS ability to counteract RT-induced ROS accumulation, antioxidant enzyme and miRNA expression and was able to increase the abundance of γ-H2AX, a biomarker of DNA damage, in RT-treated cells. Conclusions: Taken together, our data suggest the strategic role of oxidant/antioxidant balance in restraining the therapeutic efficiency of RT in RMS treatment and identify NRF2 as a new potential molecular target whose inhibition might represent a novel radiosensitizing therapeutic strategy for RMS clinical management.

NRF2 orchestrates the redox regulation induced by radiation therapy, sustaining embryonal and alveolar rhabdomyosarcoma cells radioresistance / Marampon, Francesco; Codenotti, Silvia; Megiorni, Francesca; Del Fattore, Andrea; Camero, Simona; Gravina, Giovanni Luca; Festuccia, Claudio; Musio, Daniela; De Felice, Francesca; Nardone, Valerio; Santoro, Anna Natalizia; Dominici, Carlo; Fanzani, Alessandro; Pirtoli, Luigi; Fioravanti, Antonella; Tombolini, Vincenzo; Cheleschi, Sara; Tini, Paolo. - In: JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY. - ISSN 0171-5216. - 145:4(2019), pp. 881-893. [10.1007/s00432-019-02851-0]

NRF2 orchestrates the redox regulation induced by radiation therapy, sustaining embryonal and alveolar rhabdomyosarcoma cells radioresistance

Marampon, Francesco
Primo
;
Megiorni, Francesca;Camero, Simona;Gravina, Giovanni Luca;De Felice, Francesca;Dominici, Carlo;Fioravanti, Antonella;Tombolini, Vincenzo;
2019

Abstract

Purpose: Tumor cells generally exhibit higher levels of reactive oxygen species (ROS), however, when stressed, tumor cells can undergo a process of ‘Redox Resetting’ to acquire a new redox balance with stronger antioxidant systems that enable cancer cells to become resistant to radiation therapy (RT). Here, we describe how RT affects the oxidant/antioxidant balance in human embryonal (RD) and alveolar (RH30) rhabdomyosarcoma (RMS) cell lines, investigating on the molecular mechanisms involved. Methods: Radiations were delivered using an x-6 MV photon linear accelerator and their effects were assessed by vitality and clonogenic assays. The expression of specific antioxidant-enzymes, such as Superoxide Dismutases (SODs), Catalase (CAT) and Glutathione Peroxidases 4 (GPx4), miRNAs (miR-22, -126, -210, -375, -146a, -34a) and the transcription factor NRF2 was analyzed by quantitative polymerase chain reaction (q-PCR) and western blotting. RNA interference experiments were performed to evaluate the role of NRF2. Results: Doses of RT higher than 2 Gy significantly affected RMS clonogenic ability by increasing ROS production. RMS rapidly and efficiently brought back ROS levels by up-regulating the gene expression of antioxidant enzymes, miRNAs as well as of NRF2. Silencing of NRF2 restrained the RMS ability to counteract RT-induced ROS accumulation, antioxidant enzyme and miRNA expression and was able to increase the abundance of γ-H2AX, a biomarker of DNA damage, in RT-treated cells. Conclusions: Taken together, our data suggest the strategic role of oxidant/antioxidant balance in restraining the therapeutic efficiency of RT in RMS treatment and identify NRF2 as a new potential molecular target whose inhibition might represent a novel radiosensitizing therapeutic strategy for RMS clinical management.
2019
anti-oxidant; NRF2; radioresistance; radiotherapy; reactive oxygen species; rhabdomyosarcoma; oncology; cancer research
01 Pubblicazione su rivista::01a Articolo in rivista
NRF2 orchestrates the redox regulation induced by radiation therapy, sustaining embryonal and alveolar rhabdomyosarcoma cells radioresistance / Marampon, Francesco; Codenotti, Silvia; Megiorni, Francesca; Del Fattore, Andrea; Camero, Simona; Gravina, Giovanni Luca; Festuccia, Claudio; Musio, Daniela; De Felice, Francesca; Nardone, Valerio; Santoro, Anna Natalizia; Dominici, Carlo; Fanzani, Alessandro; Pirtoli, Luigi; Fioravanti, Antonella; Tombolini, Vincenzo; Cheleschi, Sara; Tini, Paolo. - In: JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY. - ISSN 0171-5216. - 145:4(2019), pp. 881-893. [10.1007/s00432-019-02851-0]
File allegati a questo prodotto
File Dimensione Formato  
Maranpon_NRF2_2019.pdf

solo gestori archivio

Tipologia: Versione editoriale (versione pubblicata con il layout dell'editore)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 1.52 MB
Formato Adobe PDF
1.52 MB Adobe PDF   Contatta l'autore

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1261329
Citazioni
  • ???jsp.display-item.citation.pmc??? 14
  • Scopus 27
  • ???jsp.display-item.citation.isi??? 26
social impact