Inhibition of phosphorylated c-Met in rhabdomyosarcoma cell lines by a small molecule inhibitor SU11274

Abstract

Backgroundc-Met is a receptor tyrosine kinase (RTK) that is over-expressed in a variety of cancers and involved in cell growth, invasion, metastasis and angiogenesis. In this study, we investigated the role of c-Met in rhabdomyosarcoma (RMS) using its small molecule inhibitor SU11274, which has been hypothesized to be a potential therapeutic target for RMS.MethodsThe expression level of phosphorylated c-Met in RMS cell lines (RD, CW9019 and RH30) and tumor tissues was assessed by phospho-RTK array and immunohistochemistry, respectively. The inhibition effects of SU11274 on RMS cells were studied with regard to intracellular signaling, cell proliferation, cell cycle and cell migration.ResultsA high level of phosphorylated c-Met was detected in 2 alveolar RMS cell lines (CW9019 and RH30) and 14 out of 24 RMS tissue samples, whereas relatively low levels of phospho-c-Met were observed in the embryonic RMS cell line (RD). The small molecule SU11274 could significantly reduce the phosphorylation of c-Met, resulting in inhibition of cell proliferation, G1 phase arrest of cell cycle and blocking of cell migration in CW9019 and RH30 cell lines.ConclusionThese results might support the role of c-Met in the development and progression of RMS. Furthermore, the inhibitor of c-Met, SU11274, could be an effective targeting therapy reagent for RMS, especially alveolar RMS.

In vitro evaluation of the Aurora kinase inhibitor VX-680 for Hepatoblastoma

“…For SU11274, a moderate effect on cell viability was observed even though the highest concentrations were about tenfold of what is regularly used for in vitro assays [36][37][38]. This result may be due to low levels of phosphorylated Met in HB cells since there is evidence that the effectiveness of SU11274 also depends on the phosphorylation status of Met [36]. The inhibitor may also counteract cell proliferation when exogenous HGF is added, however, in cultures with serum stimulation proliferation of HB cells seems not to be driven by Met [39].…”

“…Met is highly overexpressed in a number of solid tumors, including hepatocellular carcinoma, and there is evidence that overexpression correlates with an aggressive phenotype and poor prognosis [32,34,35]. For SU11274, a moderate effect on cell viability was observed even though the highest concentrations were about tenfold of what is regularly used for in vitro assays [36][37][38]. This result may be due to low levels of phosphorylated Met in HB cells since there is evidence that the effectiveness of SU11274 also depends on the phosphorylation status of Met [36].…”

Quantitative Phosphoproteomics Analysis Reveals Broad Regulatory Role of Heparan Sulfate on Endothelial Signaling

“…Phospho-Receptor Tyrosine Kinase (p-RTK) Array Assay-Mouse p-RTK Array (R&D Systems) was employed to determine the relative level of tyrosine phosphorylation of 39 different receptor tyrosine kinases (RTK) in Ext1f/f and Ext1Ϫ/Ϫ MLECs cultured under normal serum-containing conditions, and the experiment was carried out according to the instructions provided by the manufacturer and as previously described (31). In brief, MLECs cultured in DMEM with 10% FBS was extracted with RIPA buffer (Pierce, Thermo Fisher) supplemented with phosphatase inhibitors (1:100, Sigma-Aldrich, St. Louis, MO) and protease inhibitors (1:10, Sigma-Aldrich).…”

Heme Oxygenase-1 Controls an HDAC4-miR-206 Pathway of Oxidative Stress in Rhabdomyosarcoma

“…Both Pax3-FoxO1 and Pax7-FoxO1 are more active than Pax3 and Pax7 (8) and augment expression of CXCR4, SDF1, cMET, and HGF, proteins facilitating aRMS progression (6). HGF and cMET increase RMS proliferation and invasion while reduce apoptosis, correlating with increased aggressiveness of tumors (29). Transfection of eRMS with Pax3-FoxO1 increased growth and accelerated tumor formation in mice (8,30).…”