Hyper-activation of notch3 amplifies the proliferative potential of rhabdomyosarcoma cells

Maria De Salvo, Lavinia Raimondi, Serena Vella, Laura Adesso, Roberta Ciarapica, Federica Verginelli, Antonio Pannuti, Arianna Citti, Renata Boldrini, Giuseppe M. Milano, Antonella Cacchione, Andrea Ferrari, Paola Collini, Angelo Rosolen, Gianni Bisogno, Rita Alaggio, Alessandro Inserra, Mattia Locatelli, Stefano Stifani, Isabella Screpanti & 3 others Lucio Miele, Franco Locatelli, Rossella Rota

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Rhabdomyosarcoma (RMS) is a pediatric myogenic-derived soft tissue sarcoma that includes two major histopathological subtypes: embryonal and alveolar. The majority of alveolar RMS expresses PAX3-FOXO1 fusion oncoprotein, associated with the worst prognosis. RMS cells show myogenic markers expression but are unable to terminally differentiate. The Notch signaling pathway is a master player during myogenesis, with Notch1 activation sustaining myoblast expansion and Notch3 activation inhibiting myoblast fusion and differentiation. Accordingly, Notch1 signaling is up-regulated and activated in embryonal RMS samples and supports the proliferation of tumor cells. However, it is unable to control their differentiation properties. We previously reported that Notch3 is activated in RMS cell lines, of both alveolar and embryonal subtype, and acts by inhibiting differentiation. Moreover, Notch3 depletion reduces PAX3-FOXO1 alveolar RMS tumor growth in vivo. However, whether Notch3 activation also sustains the proliferation of RMS cells remained unclear. To address this question, we forced the expression of the activated form of Notch3, Notch3IC, in the RH30 and RH41 PAX3-FOXO1-positive alveolar and in the RD embryonal RMS cell lines and studied the proliferation of these cells. We show that, in all three cell lines tested, Notch3IC over-expression stimulates in vitro cell proliferation and prevents the effects of pharmacological Notch inhibition. Furthermore, Notch3IC further increases RH30 cell growth in vivo. Interestingly, knockdown of Notch canonical ligands JAG1 or DLL1 in RMS cell lines decreases Notch3 activity and reduces cell proliferation. Finally, the expression of Notch3IC and its target gene HES1 correlates with that of the proliferative marker Ki67 in a small cohort of primary PAX-FOXO1 alveolar RMS samples. These results strongly suggest that high levels of Notch3 activation increase the proliferative potential of RMS cells.

Original languageEnglish
Article numbere96238
JournalPLoS One
Volume9
Issue number5
DOIs
Publication statusPublished - May 5 2014

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Rhabdomyosarcoma
cell proliferation
Chemical activation
Alveolar Rhabdomyosarcoma
Cells
cell lines
myoblasts
Cell Proliferation
Embryonal Rhabdomyosarcoma
Cell proliferation
Cell Line
Myoblasts
cells
Tumors
Fusion reactions
muscle development
sarcoma
Pediatrics
prognosis
Oncogene Proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Hyper-activation of notch3 amplifies the proliferative potential of rhabdomyosarcoma cells. / De Salvo, Maria; Raimondi, Lavinia; Vella, Serena; Adesso, Laura; Ciarapica, Roberta; Verginelli, Federica; Pannuti, Antonio; Citti, Arianna; Boldrini, Renata; Milano, Giuseppe M.; Cacchione, Antonella; Ferrari, Andrea; Collini, Paola; Rosolen, Angelo; Bisogno, Gianni; Alaggio, Rita; Inserra, Alessandro; Locatelli, Mattia; Stifani, Stefano; Screpanti, Isabella; Miele, Lucio; Locatelli, Franco; Rota, Rossella.

In: PLoS One, Vol. 9, No. 5, e96238, 05.05.2014.

Research output: Contribution to journalArticle

De Salvo, M, Raimondi, L, Vella, S, Adesso, L, Ciarapica, R, Verginelli, F, Pannuti, A, Citti, A, Boldrini, R, Milano, GM, Cacchione, A, Ferrari, A, Collini, P, Rosolen, A, Bisogno, G, Alaggio, R, Inserra, A, Locatelli, M, Stifani, S, Screpanti, I, Miele, L, Locatelli, F & Rota, R 2014, 'Hyper-activation of notch3 amplifies the proliferative potential of rhabdomyosarcoma cells', PLoS One, vol. 9, no. 5, e96238. https://doi.org/10.1371/journal.pone.0096238
De Salvo, Maria ; Raimondi, Lavinia ; Vella, Serena ; Adesso, Laura ; Ciarapica, Roberta ; Verginelli, Federica ; Pannuti, Antonio ; Citti, Arianna ; Boldrini, Renata ; Milano, Giuseppe M. ; Cacchione, Antonella ; Ferrari, Andrea ; Collini, Paola ; Rosolen, Angelo ; Bisogno, Gianni ; Alaggio, Rita ; Inserra, Alessandro ; Locatelli, Mattia ; Stifani, Stefano ; Screpanti, Isabella ; Miele, Lucio ; Locatelli, Franco ; Rota, Rossella. / Hyper-activation of notch3 amplifies the proliferative potential of rhabdomyosarcoma cells. In: PLoS One. 2014 ; Vol. 9, No. 5.
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abstract = "Rhabdomyosarcoma (RMS) is a pediatric myogenic-derived soft tissue sarcoma that includes two major histopathological subtypes: embryonal and alveolar. The majority of alveolar RMS expresses PAX3-FOXO1 fusion oncoprotein, associated with the worst prognosis. RMS cells show myogenic markers expression but are unable to terminally differentiate. The Notch signaling pathway is a master player during myogenesis, with Notch1 activation sustaining myoblast expansion and Notch3 activation inhibiting myoblast fusion and differentiation. Accordingly, Notch1 signaling is up-regulated and activated in embryonal RMS samples and supports the proliferation of tumor cells. However, it is unable to control their differentiation properties. We previously reported that Notch3 is activated in RMS cell lines, of both alveolar and embryonal subtype, and acts by inhibiting differentiation. Moreover, Notch3 depletion reduces PAX3-FOXO1 alveolar RMS tumor growth in vivo. However, whether Notch3 activation also sustains the proliferation of RMS cells remained unclear. To address this question, we forced the expression of the activated form of Notch3, Notch3IC, in the RH30 and RH41 PAX3-FOXO1-positive alveolar and in the RD embryonal RMS cell lines and studied the proliferation of these cells. We show that, in all three cell lines tested, Notch3IC over-expression stimulates in vitro cell proliferation and prevents the effects of pharmacological Notch inhibition. Furthermore, Notch3IC further increases RH30 cell growth in vivo. Interestingly, knockdown of Notch canonical ligands JAG1 or DLL1 in RMS cell lines decreases Notch3 activity and reduces cell proliferation. Finally, the expression of Notch3IC and its target gene HES1 correlates with that of the proliferative marker Ki67 in a small cohort of primary PAX-FOXO1 alveolar RMS samples. These results strongly suggest that high levels of Notch3 activation increase the proliferative potential of RMS cells.",
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AU - De Salvo, Maria

AU - Raimondi, Lavinia

AU - Vella, Serena

AU - Adesso, Laura

AU - Ciarapica, Roberta

AU - Verginelli, Federica

AU - Pannuti, Antonio

AU - Citti, Arianna

AU - Boldrini, Renata

AU - Milano, Giuseppe M.

AU - Cacchione, Antonella

AU - Ferrari, Andrea

AU - Collini, Paola

AU - Rosolen, Angelo

AU - Bisogno, Gianni

AU - Alaggio, Rita

AU - Inserra, Alessandro

AU - Locatelli, Mattia

AU - Stifani, Stefano

AU - Screpanti, Isabella

AU - Miele, Lucio

AU - Locatelli, Franco

AU - Rota, Rossella

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