Crizotinib-induced antitumour activity in human alveolar rhabdomyosarcoma cells is not solely dependent on ALK and MET inhibitionReport as inadecuate

Crizotinib-induced antitumour activity in human alveolar rhabdomyosarcoma cells is not solely dependent on ALK and MET inhibition - Download this document for free, or read online. Document in PDF available to download.

Journal of Experimental and Clinical Cancer Research

, 34:112

First Online: 06 October 2015Received: 11 June 2015Accepted: 29 September 2015DOI: 10.1186-s13046-015-0228-4

Cite this article as: Megiorni, F., McDowell, H.P., Camero, S. et al. J Exp Clin Cancer Res 2015 34: 112. doi:10.1186-s13046-015-0228-4


BackgroundRhabdomyosarcoma RMS is the most commonly diagnosed malignant soft tissue tumour in children and adolescents. Aberrant expression of Anaplastic Lymphoma Kinase ALK and MET gene has been implicated in the malignant progression of RMS, especially in the alveolar subtype. This observation suggests that crizotinib PF-02341066, a kinase inhibitor against ALK and MET, may have a therapeutic role in RMS, although its antitumour activity in this malignancy has not yet been studied.

MethodsRH4 and RH30 alveolar RMS ARMS cell lines were treated with crizotinib and then assessed by using proliferation, viability, migration and colony formation assays. Multiple approaches, including flow cytometry, immunofluorescence, western blotting and siRNA-based knock-down, were used in order to investigate possible molecular mechanisms linked to crizotinib activity.

ResultsIn vitro treatment with crizotinib inhibited ALK and MET proteins, as well as Insulin-like Growth Factor 1 Receptor IGF1R, with a concomitant robust dephosphorylation of AKT and ERK, two downstream kinases involved in RMS cell proliferation and survival. Exposure to crizotinib impaired cell growth, and accumulation at G2-M phase was attributed to an altered expression and activation of checkpoint regulators, such as Cyclin B1 and Cdc2. Crizotinib was able to induce apoptosis and autophagy in a dose-dependent manner, as shown by caspase-3 activation-PARP proteolytic cleavage down-regulation and by LC3 activation-p62 down-regulation, respectively. The accumulation of reactive oxygen species ROS seemed to contribute to crizotinib effects in RH4 and RH30 cells. Moreover, crizotinib-treated RH4 and RH30 cells exhibited a decreased migratory-invasive capacity and clonogenic potential.

ConclusionsThese results provide a further insight into the molecular mechanisms affected by crizotinib in ARMS cells inferring that it could be a useful therapeutic tool in ARMS cancer treatment.

KeywordsCrizotinib ALK MET Alveolar rhabdomyosarcoma G2-M arrest IGF1R Electronic supplementary materialThe online version of this article doi:10.1186-s13046-015-0228-4 contains supplementary material, which is available to authorized users.

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Author: Francesca Megiorni - Heather P. McDowell - Simona Camero - Olga Mannarino - Simona Ceccarelli - Milena Paiano - Paul D. L


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