Limits...
SRC family kinase (SFK) inhibition reduces rhabdomyosarcoma cell growth in vitro and in vivo and triggers p38 MAP kinase-mediated differentiation.

Casini N, Forte IM, Mastrogiovanni G, Pentimalli F, Angelucci A, Festuccia C, Tomei V, Ceccherini E, Di Marzo D, Schenone S, Botta M, Giordano A, Indovina P - Oncotarget (2015)

Bottom Line: SI221, which showed to be mainly effective against the SFK member YES, significantly reduced cell viability and induced apoptosis, without affecting non-tumor cells, such as primary human skin fibroblasts and differentiated C2C12 cells.Moreover, SI221 decreased in vitro cell migration and invasion and reduced tumor growth in a RMS xenograft model.Overall, our findings suggest that SFK inhibition, besides reducing RMS cell growth and invasive potential, could also represent a differentiation therapeutic strategy for RMS.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Surgery and Neuroscience, University of Siena and Istituto Toscano Tumori (ITT), Siena, Italy.

ABSTRACT
Recent data suggest that SRC family kinases (SFKs) could represent potential therapeutic targets for rhabdomyosarcoma (RMS), the most common soft-tissue sarcoma in children. Here, we assessed the effect of a recently developed selective SFK inhibitor (a pyrazolo[3,4-d]pyrimidine derivative, called SI221) on RMS cell lines. SI221, which showed to be mainly effective against the SFK member YES, significantly reduced cell viability and induced apoptosis, without affecting non-tumor cells, such as primary human skin fibroblasts and differentiated C2C12 cells. Moreover, SI221 decreased in vitro cell migration and invasion and reduced tumor growth in a RMS xenograft model. SFK inhibition also induced muscle differentiation in RMS cells by affecting the NOTCH3 receptor-p38 mitogen-activated protein kinase (MAPK) axis, which regulates the balance between proliferation and differentiation. Overall, our findings suggest that SFK inhibition, besides reducing RMS cell growth and invasive potential, could also represent a differentiation therapeutic strategy for RMS.

No MeSH data available.


Related in: MedlinePlus

Apoptosis induction in RMS cell lines treated with SI221(A) A representative FACS analysis to investigate apoptosis by cell staining with annexin V-FITC and propidium iodide (PI) of RD and RH30 cells 72 hours after treatment with SI221 or DMSO, as a control. The table reports the values relative to early apoptosis (annexin V positive and PI negative), late apoptosis (annexin V positive and PI positive) and total apoptosis. (B) Histograms showing caspase-3 activity in RD and RH30 cell lines 72 hours after treatment with SI221 or DMSO, as a control. Caspase-3 activity is expressed as pmol pNA/μg protein × time (hours). The reported values represent the means and standard deviations of three independent experiments. Statistically significant differences were evaluated by Student t test and indicated with *: significant (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4494948&req=5

Figure 2: Apoptosis induction in RMS cell lines treated with SI221(A) A representative FACS analysis to investigate apoptosis by cell staining with annexin V-FITC and propidium iodide (PI) of RD and RH30 cells 72 hours after treatment with SI221 or DMSO, as a control. The table reports the values relative to early apoptosis (annexin V positive and PI negative), late apoptosis (annexin V positive and PI positive) and total apoptosis. (B) Histograms showing caspase-3 activity in RD and RH30 cell lines 72 hours after treatment with SI221 or DMSO, as a control. Caspase-3 activity is expressed as pmol pNA/μg protein × time (hours). The reported values represent the means and standard deviations of three independent experiments. Statistically significant differences were evaluated by Student t test and indicated with *: significant (P < 0.05).

Mentions: To evaluate SI221 ability to induce apoptosis in RD and RH30 cell lines, we analyzed cell staining with annexin V-FITC and PI by FACS 72 hours after treatment with SI221 at its IC50 values. Analyses of early and late apoptosis showed that SI221 was able to induce apoptosis in both the cell lines (Figure 2A). Consistently, we observed a significant increase in caspase-3 activity in RD and RH30 cells 72 hours after treatment with SI221 at its IC50 values (Figure 2B).


SRC family kinase (SFK) inhibition reduces rhabdomyosarcoma cell growth in vitro and in vivo and triggers p38 MAP kinase-mediated differentiation.

Casini N, Forte IM, Mastrogiovanni G, Pentimalli F, Angelucci A, Festuccia C, Tomei V, Ceccherini E, Di Marzo D, Schenone S, Botta M, Giordano A, Indovina P - Oncotarget (2015)

Apoptosis induction in RMS cell lines treated with SI221(A) A representative FACS analysis to investigate apoptosis by cell staining with annexin V-FITC and propidium iodide (PI) of RD and RH30 cells 72 hours after treatment with SI221 or DMSO, as a control. The table reports the values relative to early apoptosis (annexin V positive and PI negative), late apoptosis (annexin V positive and PI positive) and total apoptosis. (B) Histograms showing caspase-3 activity in RD and RH30 cell lines 72 hours after treatment with SI221 or DMSO, as a control. Caspase-3 activity is expressed as pmol pNA/μg protein × time (hours). The reported values represent the means and standard deviations of three independent experiments. Statistically significant differences were evaluated by Student t test and indicated with *: significant (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494948&req=5

Figure 2: Apoptosis induction in RMS cell lines treated with SI221(A) A representative FACS analysis to investigate apoptosis by cell staining with annexin V-FITC and propidium iodide (PI) of RD and RH30 cells 72 hours after treatment with SI221 or DMSO, as a control. The table reports the values relative to early apoptosis (annexin V positive and PI negative), late apoptosis (annexin V positive and PI positive) and total apoptosis. (B) Histograms showing caspase-3 activity in RD and RH30 cell lines 72 hours after treatment with SI221 or DMSO, as a control. Caspase-3 activity is expressed as pmol pNA/μg protein × time (hours). The reported values represent the means and standard deviations of three independent experiments. Statistically significant differences were evaluated by Student t test and indicated with *: significant (P < 0.05).
Mentions: To evaluate SI221 ability to induce apoptosis in RD and RH30 cell lines, we analyzed cell staining with annexin V-FITC and PI by FACS 72 hours after treatment with SI221 at its IC50 values. Analyses of early and late apoptosis showed that SI221 was able to induce apoptosis in both the cell lines (Figure 2A). Consistently, we observed a significant increase in caspase-3 activity in RD and RH30 cells 72 hours after treatment with SI221 at its IC50 values (Figure 2B).

Bottom Line: SI221, which showed to be mainly effective against the SFK member YES, significantly reduced cell viability and induced apoptosis, without affecting non-tumor cells, such as primary human skin fibroblasts and differentiated C2C12 cells.Moreover, SI221 decreased in vitro cell migration and invasion and reduced tumor growth in a RMS xenograft model.Overall, our findings suggest that SFK inhibition, besides reducing RMS cell growth and invasive potential, could also represent a differentiation therapeutic strategy for RMS.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Surgery and Neuroscience, University of Siena and Istituto Toscano Tumori (ITT), Siena, Italy.

ABSTRACT
Recent data suggest that SRC family kinases (SFKs) could represent potential therapeutic targets for rhabdomyosarcoma (RMS), the most common soft-tissue sarcoma in children. Here, we assessed the effect of a recently developed selective SFK inhibitor (a pyrazolo[3,4-d]pyrimidine derivative, called SI221) on RMS cell lines. SI221, which showed to be mainly effective against the SFK member YES, significantly reduced cell viability and induced apoptosis, without affecting non-tumor cells, such as primary human skin fibroblasts and differentiated C2C12 cells. Moreover, SI221 decreased in vitro cell migration and invasion and reduced tumor growth in a RMS xenograft model. SFK inhibition also induced muscle differentiation in RMS cells by affecting the NOTCH3 receptor-p38 mitogen-activated protein kinase (MAPK) axis, which regulates the balance between proliferation and differentiation. Overall, our findings suggest that SFK inhibition, besides reducing RMS cell growth and invasive potential, could also represent a differentiation therapeutic strategy for RMS.

No MeSH data available.


Related in: MedlinePlus