Abstract
All-trans retinoic acid is a potent promoter of cellular differentiation processes, which is used in cancer therapy. Glioblastoma spheroid cultures are enriched in tumor-initiating cells, and provide a model to test new treatment options in vitro. We investigated the molecular mechanisms of response to exposure to differentiation-promoting conditions in such cultures. Microarray analyses of five independent cultures showed that after induction of differentiation, inhibitors of transforming growth factor-β/bone morphogenetic protein, Wnt/β-catenin and IGF signaling were upregulated, whereas expression of several microRNAs decreased, particularly that of the miR-17-92 cluster. In primary astrocytic gliomas (n=82), expression of several members of miR-17-92 was significantly higher relative to those of normal brain (n=8) and significantly increased with tumor grade progression (P<0.05). A high-level amplification of the miR-17-92 locus was detected in one glioblastoma specimen. Transfection of inhibitors of miR-17-92 induced increased apoptosis and decreased cell proliferation in glioblastoma spheroids. Mir-17-92 inhibition was also associated with increased messenger RNA (mRNA) and/or protein expression of CDKN1A, E2F1, PTEN and CTGF. The CTGF gene was shown to be a target of miR-17-92 in glioblastoma spheroids by luciferase reporter assays. Our results suggest that miR-17-92 and its target CTGF mediate effects of differentiation-promoting treatment on glioblastoma cells through multiple regulatory pathways.
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Acknowledgements
We thank F Engel, M Zapatka, S Anders and W Huber for bioinformatic support. We also thank S Hofmann for support with array comparative genomic hybridization, A Korshunov for support with specimen analysis and B Malzkorn for sharing data. This study was supported by Grants 01GS0883, 01GS0884 and 01GS0886 of the German Bundesministerium für Bildung und Forschung, by Grant MRTN-CT-2006-035733 from the European Union FP6 Marie Curie Research Training Network; by a Grant from the Tumorzentrum Heidelberg-Mannheim; by the Sibylle Assmus Foundation; and by the Verein zur Förderung der Krebsforschung e.V.
Data Deposition: Gene expression data are available at Gene Expression Omnibus accession no. GSE17227 (http://www.ncbi.nlm.nih.gov/geo/).
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Ernst, A., Campos, B., Meier, J. et al. De-repression of CTGF via the miR-17-92 cluster upon differentiation of human glioblastoma spheroid cultures. Oncogene 29, 3411–3422 (2010). https://doi.org/10.1038/onc.2010.83
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DOI: https://doi.org/10.1038/onc.2010.83
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