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Increased miR-155-5p and reduced miR-148a-3p contribute to the suppression of osteosarcoma cell death

Oncogene volume 35, pages 5282–5294 (2016)Cite this article

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Abstract

Osteosarcoma (OS) is the most common cancer of bone and the 5th leading cause of cancer-related death in young adults. Currently, 5-year survival rates have plateaued at ~70% for patients with localized disease. Those with disseminated disease have an ~20% 5-year survival. An improved understanding of the molecular genetics of OS may yield new approaches to improve outcomes for OS patients. To this end, we applied murine models that replicate human OS to identify and understand dysregulated microRNAs (miRNAs) in OS. miRNA expression patterns were profiled in murine primary osteoblasts, osteoblast cultures and primary OS cell cultures (from primary and paired metastatic locations) isolated from two genetically engineered murine models of OS. The differentially expressed miRNA were further assessed by a cross-species comparison with human osteoblasts and OS cultures. We identified miR-155-5p and miR-148a-3p as deregulated in OS. miR-155-5p suppression or miR-148a-3p overexpression potently reduced proliferation and induced apoptosis in OS cells, yet strikingly, did not impact normal osteoblasts. To define how these miRNAs regulated OS cell fate, we used an integrated computational approach to identify putative candidate targets and then correlated these with the cell biological impact. Although we could not resolve the mechanism through which miR-148a-3p impacts OS, we identified that miR-155-5p overexpression suppressed its target Ripk1 (receptor (TNFRSF)-interacting serine–threonine kinase 1) expression, and miR-155-5p inhibition elevated Ripk1 levels. Ripk1 is directly involved in apoptosis/necroptosis. In OS cells, small interfering RNA against Ripk1 prevented cell death induced by the sequestration of miR-155-5p. Collectively, we show that miR-148a-3p and miR-155-5p are species-conserved deregulated miRNA in OS. Modulation of these miRNAs was specifically toxic to tumor cells but not normal osteoblasts, raising the possibility that these may be tractable targets for miRNA-based therapies for OS.

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Acknowledgements

We thank A Pfeiffer (University of Bonn) for providing the miR-155-5p sponge, control sponge and miR-155-5p overexpression vector. We thank A Gupte for technical assistance; C Hawkins for discussion and comments; the BioResource Facility (St Vincent’s Hospital) for housing and care of experimental animals, and M Thomson (SVI Flow Cytometry Facility) for help with FACS analysis. This work was supported by grants from the National Health and Medical Research Council (NHMRC), Australia (CW and TJM), Cancer Council of Victoria (CW and EB); Cure Cancer Australia Foundation (EB); 5point foundation (EB); Zig Inge Foundation (CW); NHMRC Career Development Award (CW); NHMRC Dora Lush postgraduate scholarship (SB); in part by the Victorian State Government OIS Program (to St. Vincent’s Institute); CW was the Phillip Desbrow Senior Research Fellow of the Leukaemia Foundation.

Author contributions

SB, CRW, EB, JL, AMC and AJN performed the experiments, analyzed and interpreted data; SB, CRW, EB, JL, TJM and LEP provided the intellectual input and conceptual advice; AZ and JL provided the samples and reagents; SB and CRW wrote the manuscript; all authors reviewed and edited the manuscript.

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  1. E K Baker and C R Walkley: These authors contributed equally to this work.

Authors and Affiliations

  1. St Vincent’s Institute of Medical Research and Department of Medicine, St Vincent’s Hospital, University of Melbourne, Fitzroy, Victoria, Australia

    S Bhattacharya, A M Chalk, A J M Ng, T J Martin, L E Purton, E K Baker & C R Walkley

  2. Myeloma Research Laboratory, School of Medicine, Faculty of Health Sciences, University of Adelaide, Adelaide, South Australia, Australia

    A C Zannettino

  3. Cancer Theme, South Australian Health and Medical Research Institute, Adelaide, South Australia, Australia

    A C Zannettino

  4. ACRF Rational Drug Discovery Centre, St Vincent’s Institute of Medical Research, Fitzroy, Victoria, Australia

    L E Purton & C R Walkley

  5. Department of Genetics and Yale Stem Cell Center, Yale University, New Haven, CT, USA

    J Lu

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  1. S Bhattacharya
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  2. A M Chalk
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Correspondence to C R Walkley.

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Bhattacharya, S., Chalk, A., Ng, A. et al. Increased miR-155-5p and reduced miR-148a-3p contribute to the suppression of osteosarcoma cell death. Oncogene 35, 5282–5294 (2016). https://doi.org/10.1038/onc.2016.68

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