MURC/cavin-4 is co-expressed with Caveolin-3 in rhabdomyosarcoma tumors and its silencing prevents myogenic differentiation in the human embryonal RD cell line

Fiorella Faggi, Silvia Codenotti, Pietro Luigi Poliani, Manuela Cominelli, Nicola Chiarelli, Marina Colombi, Marika Vezzoli, Eugenio Monti, Federica Bono, Giovanni Tulipano, Chiara Fiorentini, Alessandra Zanola, Harriet P. Lo, Robert G. Parton, Charles Keller, Alessandro Fanzani

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Abstract

The purpose of this study was to investigate whether MURC/cavin-4, a plasma membrane and Z-line associated protein exhibiting an overlapping distribution with Caveolin-3 (Cav-3) in heart and muscle tissues, may be expressed and play a role in rhabdomyosarcoma (RMS), an aggressive myogenic tumor affecting childhood. We found MURC/cavin-4 to be expressed, often concurrently with Cav-3, in mouse and human RMS, as demonstrated through in silico analysis of gene datasets and immunohistochemical analysis of tumor samples. In vitro expression studies carried out using human cell lines and primary mouse tumor cultures showed that expression levels of both MURC/cavin-4 and Cav-3, while being low or undetectable during cell proliferation, became robustly increased during myogenic differentiation, as detected via semi-quantitative RT-PCR and immunoblotting analysis. Furthermore, confocal microscopy analysis performed on human RD and RH30 cell lines confirmed that MURC/cavin-4 mostly marks differentiated cell elements, colocalizing at the cell surface with Cav-3 and labeling myosin heavy chain (MHC) expressing cells. Finally, MURC/cavin-4 silencing prevented the differentiation in the RD cell line, leading to morphological cell impairment characterized by depletion of myogenin, Cav-3 and MHC protein levels. Overall, our data suggest that MURC/cavin-4, especially in combination with Cav-3, may play a consistent role in the differentiation process of RMS.

Original languageEnglish (US)
Article numbere0130287
JournalPLoS One
Volume10
Issue number6
DOIs
StatePublished - Jun 18 2015

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Caveolin 3
Rhabdomyosarcoma
Tumors
Cells
cell lines
Cell Line
neoplasms
myosin heavy chains
Neoplasms
Myosin Heavy Chains
cells
human cell lines
mice
immunoblotting
muscle tissues
childhood
Myogenin
cell proliferation
plasma membrane
proteins

ASJC Scopus subject areas

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

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MURC/cavin-4 is co-expressed with Caveolin-3 in rhabdomyosarcoma tumors and its silencing prevents myogenic differentiation in the human embryonal RD cell line. / Faggi, Fiorella; Codenotti, Silvia; Poliani, Pietro Luigi; Cominelli, Manuela; Chiarelli, Nicola; Colombi, Marina; Vezzoli, Marika; Monti, Eugenio; Bono, Federica; Tulipano, Giovanni; Fiorentini, Chiara; Zanola, Alessandra; Lo, Harriet P.; Parton, Robert G.; Keller, Charles; Fanzani, Alessandro.

In: PLoS One, Vol. 10, No. 6, e0130287, 18.06.2015.

Research output: Contribution to journalArticle

Faggi, F, Codenotti, S, Poliani, PL, Cominelli, M, Chiarelli, N, Colombi, M, Vezzoli, M, Monti, E, Bono, F, Tulipano, G, Fiorentini, C, Zanola, A, Lo, HP, Parton, RG, Keller, C & Fanzani, A 2015, 'MURC/cavin-4 is co-expressed with Caveolin-3 in rhabdomyosarcoma tumors and its silencing prevents myogenic differentiation in the human embryonal RD cell line', PLoS One, vol. 10, no. 6, e0130287. https://doi.org/10.1371/journal.pone.0130287
Faggi, Fiorella ; Codenotti, Silvia ; Poliani, Pietro Luigi ; Cominelli, Manuela ; Chiarelli, Nicola ; Colombi, Marina ; Vezzoli, Marika ; Monti, Eugenio ; Bono, Federica ; Tulipano, Giovanni ; Fiorentini, Chiara ; Zanola, Alessandra ; Lo, Harriet P. ; Parton, Robert G. ; Keller, Charles ; Fanzani, Alessandro. / MURC/cavin-4 is co-expressed with Caveolin-3 in rhabdomyosarcoma tumors and its silencing prevents myogenic differentiation in the human embryonal RD cell line. In: PLoS One. 2015 ; Vol. 10, No. 6.
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abstract = "The purpose of this study was to investigate whether MURC/cavin-4, a plasma membrane and Z-line associated protein exhibiting an overlapping distribution with Caveolin-3 (Cav-3) in heart and muscle tissues, may be expressed and play a role in rhabdomyosarcoma (RMS), an aggressive myogenic tumor affecting childhood. We found MURC/cavin-4 to be expressed, often concurrently with Cav-3, in mouse and human RMS, as demonstrated through in silico analysis of gene datasets and immunohistochemical analysis of tumor samples. In vitro expression studies carried out using human cell lines and primary mouse tumor cultures showed that expression levels of both MURC/cavin-4 and Cav-3, while being low or undetectable during cell proliferation, became robustly increased during myogenic differentiation, as detected via semi-quantitative RT-PCR and immunoblotting analysis. Furthermore, confocal microscopy analysis performed on human RD and RH30 cell lines confirmed that MURC/cavin-4 mostly marks differentiated cell elements, colocalizing at the cell surface with Cav-3 and labeling myosin heavy chain (MHC) expressing cells. Finally, MURC/cavin-4 silencing prevented the differentiation in the RD cell line, leading to morphological cell impairment characterized by depletion of myogenin, Cav-3 and MHC protein levels. Overall, our data suggest that MURC/cavin-4, especially in combination with Cav-3, may play a consistent role in the differentiation process of RMS.",
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AU - Faggi, Fiorella

AU - Codenotti, Silvia

AU - Poliani, Pietro Luigi

AU - Cominelli, Manuela

AU - Chiarelli, Nicola

AU - Colombi, Marina

AU - Vezzoli, Marika

AU - Monti, Eugenio

AU - Bono, Federica

AU - Tulipano, Giovanni

AU - Fiorentini, Chiara

AU - Zanola, Alessandra

AU - Lo, Harriet P.

AU - Parton, Robert G.

AU - Keller, Charles

AU - Fanzani, Alessandro

PY - 2015/6/18

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