Article

Reduced adenosine-to-inosine miR-455-5p editing promotes melanoma growth and metastasis

Received:
Accepted:
Published online:

Abstract

Although recent studies have shown that adenosine-to-inosine (A-to-I) RNA editing occurs in microRNAs (miRNAs), its effects on tumour growth and metastasis are not well understood. We present evidence of CREB-mediated low expression of ADAR1 in metastatic melanoma cell lines and tumour specimens. Re-expression of ADAR1 resulted in the suppression of melanoma growth and metastasis in vivo. Consequently, we identified three miRNAs undergoing A-to-I editing in the weakly metastatic melanoma but not in strongly metastatic cell lines. One of these miRNAs, miR-455-5p, has two A-to-I RNA-editing sites. The biological function of edited miR-455-5p is different from that of the unedited form, as it recognizes a different set of genes. Indeed, wild-type miR-455-5p promotes melanoma metastasis through inhibition of the tumour suppressor gene CPEB1. Moreover, wild-type miR-455 enhances melanoma growth and metastasis in vivo, whereas the edited form inhibits these features. These results demonstrate a previously unrecognized role for RNA editing in melanoma progression.

  • Subscribe to Nature Cell Biology for full access:

    $59

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    , , & Cancer statistics, 2014. CA Cancer J. Clin. 64, 9–29 (2014).

  2. 2.

    & Melanoma. N. Engl. J. Med. 355, 51–65 (2006).

  3. 3.

    et al. Dominant-negative CREB inhibits tumor growth and metastasis of human melanoma cells. Oncogene 15, 2069–2075 (1997).

  4. 4.

    , , , & Driving transcriptional regulators in melanoma metastasis. Cancer Metastasis Rev. 31, 621–632 (2012).

  5. 5.

    , , , & CREB and its associated proteins act as survival factors for human melanoma cells. J. Biol. Chem. 273, 24884–24890 (1998).

  6. 6.

    & Transcriptional control of the melanoma malignant phenotype. Cancer Biol. Ther. 7, 997–1003 (2008).

  7. 7.

    , , & Platelet-activating factor mediates MMP-2 expression and activation via phosphorylation of cAMP-response element-binding protein and contributes to melanoma metastasis. J. Biol. Chem. 281, 2911–2922 (2006).

  8. 8.

    et al. Silencing cAMP-response element-binding protein (CREB) identifies CYR61 as a tumor suppressor gene in melanoma. J. Biol. Chem. 284, 26194–26206 (2009).

  9. 9.

    et al. Microphthalmia gene product as a signal transducer in cAMP-induced differentiation of melanocytes. J. Cell Biol. 142, 827–835 (1998).

  10. 10.

    et al. CREB inhibits AP-2α expression to regulate the malignant phenotype of melanoma. PLoS ONE 5, e12452 (2010).

  11. 11.

    Unbalanced alternative splicing and its significance in cancer. Bioessays 28, 378–386 (2006).

  12. 12.

    & Alternative pre-mRNA splicing regulation in cancer: pathways and programs unhinged. Genes Dev. 24, 2343–2364 (2010).

  13. 13.

    & A-to-I RNA editing and cancer: from pathology to basic science. RNA Biol. 5, 135–139 (2008).

  14. 14.

    et al. Systematic identification of abundant A-to-I editing sites in the human transcriptome. Nat. Biotechnol. 22, 1001–1005 (2004).

  15. 15.

    , , , & RNA editing and alternative splicing: the importance of co-transcriptional coordination. EMBO Rep. 7, 303–307 (2006).

  16. 16.

    , & Origins and evolution of ADAR-mediated RNA editing. IUBMB Life 61, 572–578 (2009).

  17. 17.

    RNA editing by adenosine deaminases that act on RNA. Annu. Rev. Biochem. 71, 817–846 (2002).

  18. 18.

    , & The many roles of an RNA editor. Nat. Rev. Genet. 2, 869–878 (2001).

  19. 19.

    , & A-to-I RNA editing: recent news and residual mysteries. J. Biol. Chem. 278, 1391–1394 (2003).

  20. 20.

    & Editing of neurotransmitter receptor and ion channel RNAs in the nervous system. Curr. Top. Microbiol. Immunol. 353, 61–90 (2012).

  21. 21.

    & Modulation of microRNA expression and function by ADARs. Curr. Top. Microbiol. Immunol. 353, 91–109 (2012).

  22. 22.

    & Adenosine-to-inosine RNA editing. Wiley Interdiscip. Rev. Syst. Biol. Med. 1, 202–209 (2009).

  23. 23.

    et al. Editing independent effects of ADARs on the miRNA/siRNA pathways. EMBO J. 28, 3145–3156 (2009).

  24. 24.

    et al. MicroRNA-mediated loss of ADAR1 in metastatic melanoma promotes tumor growth. J. Clin. Invest. 123, 2703–2718 (2013).

  25. 25.

    et al. Systematic identification of edited microRNAs in the human brain. Genome Res. 22, 1533–1540 (2012).

  26. 26.

    et al. A neuronal isoform of CPEB regulates local protein synthesis and stabilizes synapse-specific long-term facilitation in aplysia. Cell 115, 893–904 (2003).

  27. 27.

    et al. Over-expression of Aurora-A targets cytoplasmic polyadenylation element binding protein and promotes mRNA polyadenylation of Cdk1 and cyclin B1. Genes Cells 10, 627–638 (2005).

  28. 28.

    , , , & RNA editing of the microRNA-151 precursor blocks cleavage by the Dicer-TRBP complex. EMBO Rep. 8, 763–769 (2007).

  29. 29.

    et al. ADAR1 forms a complex with Dicer to promote MicroRNA processing and RNA-induced gene silencing. Cell 153, 575–589 (2013).

  30. 30.

    et al. Modulation of microRNA processing and expression through RNA editing by ADAR deaminases. Nat. Struct. Mol. Biol. 13, 13–21 (2006).

  31. 31.

    et al. CPEB1, a novel gene silenced in gastric cancer: a Drosophila approach. Gut 61, 1115–1123 (2012).

  32. 32.

    et al. Cytoplasmic polyadenylation element binding protein is a conserved target of tumor suppressor HRPT2/CDC73. Cell Death Differ. 17, 1551–1565 (2010).

  33. 33.

    et al. Evolutionarily conserved human targets of adenosine to inosine RNA editing. Nucleic Acids Res. 33, 1162–1168 (2005).

  34. 34.

    et al. Human BLCAP transcript: new editing events in normal and cancerous tissues. Int. J. Cancer 127, 127–137 (2010).

  35. 35.

    , , & A mammalian reporter system for fast and quantitative detection of intracellular A-to-I RNA editing levels. Anal. Biochem. 399, 230–236 (2010).

  36. 36.

    & Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25, 1754–1760 (2009).

  37. 37.

    et al. Therapeutic EphA2 gene targeting using neutral liposomal small interfering RNA delivery. Cancer Res. 65, 6910–6918 (2005).

  38. 38.

    et al. Targeting melanoma growth and metastasis with systemic delivery of liposome-incorporated protease-activated receptor-1 small interfering RNA. Cancer Res. 68, 9078–9086 (2008).

Download references

Acknowledgements

We thank S. Maas from NIH for providing the ADAR1 A-to-I RNA-editing hairpin loop luciferase reporter plasmid. We thank W. Choi for her help with the miRNA microarray. We thank R. Rajesha for his technical help and support. These studies are supported by SINF, an MDACC grant and NIH Skin Cancer SPORE p50 CA093459.

Author information

Author notes

    • Einav Shoshan
    •  & Aaron K. Mobley

    These authors contributed equally to this work.

Affiliations

  1. Department of Cancer Biology, Unit 0173, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030, USA

    • Einav Shoshan
    • , Aaron K. Mobley
    • , Russell R. Braeuer
    • , Takafumi Kamiya
    • , Li Huang
    • , Mayra E. Vasquez
    • , Ho Jeong Lee
    • , Sun Jin Kim
    • , Guermarie Velazquez-Torres
    • , Anil K. Sood
    • , Isaiah J. Fidler
    •  & Menashe Bar-Eli
  2. The University of Texas Health Science Center at Houston, 1825 Pressler Street, Houston, Texas 77030, USA

    • Ahmad Salameh
  3. Department of Gynecologic Oncology, Unit 1362, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030, USA

    • Cristina Ivan
    •  & Anil K. Sood
  4. Canada’s Michael Smith Cancer Agency, Vancouver, British Columbia V5Z 4S6, Canada

    • Ka Ming Nip
    • , Kelsey Zhu
    • , Denise Brooks
    • , Steven J. M. Jones
    • , Inanc Birol
    •  & A. Gordon Robertson
  5. Institute of Personalized Cancer Therapy, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030, USA

    • Maribel Mosqueda
    • , Yu-ye Wen
    •  & Agda Karina Eterovic
  6. Department of Melanoma Medical Oncology, Unit 0430, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030, USA

    • Patrick Hwu
  7. Department of Surgical Oncology, Unit 1484, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030, USA

    • Jeffrey E. Gershenwald
  8. Department of Experimental Therapeutics, Unit 1950, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030, USA

    • George A. Calin
  9. Ella Institute of Melanoma, Sheba Medical Center, Ramat-Gan 52621, Israel

    • Gal Markel
  10. Clinical Microbiology and Immunology, Sackler Faculty of Medicine, Tel Aviv University, Ramat Aviv 69978, Israel

    • Gal Markel

Authors

  1. Search for Einav Shoshan in:

  2. Search for Aaron K. Mobley in:

  3. Search for Russell R. Braeuer in:

  4. Search for Takafumi Kamiya in:

  5. Search for Li Huang in:

  6. Search for Mayra E. Vasquez in:

  7. Search for Ahmad Salameh in:

  8. Search for Ho Jeong Lee in:

  9. Search for Sun Jin Kim in:

  10. Search for Cristina Ivan in:

  11. Search for Guermarie Velazquez-Torres in:

  12. Search for Ka Ming Nip in:

  13. Search for Kelsey Zhu in:

  14. Search for Denise Brooks in:

  15. Search for Steven J. M. Jones in:

  16. Search for Inanc Birol in:

  17. Search for Maribel Mosqueda in:

  18. Search for Yu-ye Wen in:

  19. Search for Agda Karina Eterovic in:

  20. Search for Anil K. Sood in:

  21. Search for Patrick Hwu in:

  22. Search for Jeffrey E. Gershenwald in:

  23. Search for A. Gordon Robertson in:

  24. Search for George A. Calin in:

  25. Search for Gal Markel in:

  26. Search for Isaiah J. Fidler in:

  27. Search for Menashe Bar-Eli in:

Contributions

M.B-E. conceived and supervised the project. E.S. and A.K.M. designed and carried out experiments and analysed most of the data. R.R.B., T.K., L.H., M.E.V., G.V-T. and A.S. carried out experiments and analysed data. H.J.L., S.J.K. and I.J.F. helped with the spontaneous animal model. C.I., A.G.R., K.M.N., K.Z., D.B., S.J.M.J., I.B., M.M., Y-y.W., A.K.E., P.H. and J.E.G. carried out sequencing and analysis. A.K.S., G.A.C. and G.M. helped with miRNA analysis.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Menashe Bar-Eli.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Excel files

  1. 1.

    Supplementary Table 1

    Supplementary Information

  2. 2.

    Supplementary Table 3

    Supplementary Information