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Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3

  • Authors:
    • Shiori Kubota
    • Mitsuru Chiba
    • Miki Watanabe
    • Maki Sakamoto
    • Narumi Watanabe
  • View Affiliations / Copyright

    Affiliations: Department of Medical Technology, Hirosaki University School of Health Sciences, Hirosaki, Aomori 036-8564, Japan, Department of Biomedical Sciences, Division of Medical Life Sciences, Hirosaki University Graduate School of Health Sciences, Hirosaki, Aomori 036-8564, Japan
    Copyright: © Kubota et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].
  • Pages: 67-73
    |
    Published online on: November 13, 2014
       https://doi.org/10.3892/or.2014.3605
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Abstract

A recent study demonstrated that intracellular small/microRNAs are released from cells, and some of these extracellular RNAs are embedded in vesicles, such as ceramide-rich exosomes, on lipid-bilayer membranes. In the present study, we examined the effects of sphingomyelin phosphodiesterase 3 (SMPD3), which generates ceramide from sphingomyelin, on the release of small/microRNAs from intracellular to extracellular spaces. In these experiments, SW480 human colorectal and HuH-7 human hepatocellular cancer cells were cultured for 48 h in serum-free media. Culture supernatants were then collected, and floating cells and debris were removed by centrifugation and filtration through a 0.22-µm filter. Extracellular small RNAs in purified culture supernatants were stable for 4 weeks at room temperature, after 20 freeze-thaw cycles and exposure to pH 2.0, and were resistant to ribonuclease A degradation. Amino acid sequence analyses of SMPD3 showed high homology between mammals, indicating evolutionary conservation. Therefore, to investigate the mechanisms of cellular small/microRNA export, SW480 and HuH-7 cells were treated with the SMPD3 inhibitor GW4869 in serum-free media. Culture supernatants were collected for microarray and/or reverse transcription quantitative polymerase chain reaction (RT-qPCR) experiments. The number of microRNAs in culture supernatants was decreased following treatment with GW4869. Among these, extracellular and intracellular miR-638 were dose-dependently decreased and increased, respectively. These data suggest that SMPD3 plays an important role in the release of microRNAs into extracellular spaces.
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Copy and paste a formatted citation
Spandidos Publications style
Kubota S, Chiba M, Watanabe M, Sakamoto M and Watanabe N: Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3. Oncol Rep 33: 67-73, 2015.
APA
Kubota, S., Chiba, M., Watanabe, M., Sakamoto, M., & Watanabe, N. (2015). Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3. Oncology Reports, 33, 67-73. https://doi.org/10.3892/or.2014.3605
MLA
Kubota, S., Chiba, M., Watanabe, M., Sakamoto, M., Watanabe, N."Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3". Oncology Reports 33.1 (2015): 67-73.
Chicago
Kubota, S., Chiba, M., Watanabe, M., Sakamoto, M., Watanabe, N."Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3". Oncology Reports 33, no. 1 (2015): 67-73. https://doi.org/10.3892/or.2014.3605
Copy and paste a formatted citation
x
Spandidos Publications style
Kubota S, Chiba M, Watanabe M, Sakamoto M and Watanabe N: Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3. Oncol Rep 33: 67-73, 2015.
APA
Kubota, S., Chiba, M., Watanabe, M., Sakamoto, M., & Watanabe, N. (2015). Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3. Oncology Reports, 33, 67-73. https://doi.org/10.3892/or.2014.3605
MLA
Kubota, S., Chiba, M., Watanabe, M., Sakamoto, M., Watanabe, N."Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3". Oncology Reports 33.1 (2015): 67-73.
Chicago
Kubota, S., Chiba, M., Watanabe, M., Sakamoto, M., Watanabe, N."Secretion of small/microRNAs including miR-638 into extracellular spaces by sphingomyelin phosphodiesterase 3". Oncology Reports 33, no. 1 (2015): 67-73. https://doi.org/10.3892/or.2014.3605
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