Journal of Neuro-Oncology

, Volume 112, Issue 1, pp 27–37 | Cite as

miRNA-mediated tumor specific delivery of TRAIL reduced glioma growth

  • Yongli BoEmail author
  • Guocai Guo
  • Weicheng Yao
Laboratory Investigation


As an aggressive cancer with high morbidity, malignant glioma always has a poor prognosis even after surgery, chemotherapy and radiotherapy. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) shows a strong apoptosis-inducing effect on a variety of cancer cells including glioma. However so far, TRAIL delivery mediated by adenoviral vectors lacks tumor specificity and thus has cytotoxicity to normal cells. To improve the tumor-specificity of adenovirus-mediated TRAIL delivery, we utilized miR-124, miR-128, miR-146b and miR-218 to restrict its expression to within glioma cells. qPCR assay showed that expression of these four miRNAs was greatly downregulated in glioma in comparison with normal brain tissue. Luciferase reporter assay confirmed that miR-124, miR-128, miR-146b and miR-218 conferred exogenous gene expression with glioma-specificity. By inserting miRNA response elements (MREs) of these miRNAs into the downstream of TRAIL on adenoviral vectors, TRAIL was highly expressed in glioma cells, but not in normal brain cells. Cell viability and immunoblotting assays and FACS analysis showed that cytotoxicity and apoptosis elicited by TRAIL was only observed in glioma cells, rather than normal brain cells. Animal experiments also showed that MREs-regulated TRAIL delivery reduced the growth of glioma xenograft. In this study, we proved that miRNA-mediated tumor specific delivery of TRAIL was able to inhibit the survival of glioma cells and reduce the growth of glioma in vivo.


Glioma Adenovirus miRNA Specificity TRAIL 



We appreciated generous providing of adenoviruses by Dr. Zhao in General Hospital of Chengdu Military Area Command of Chinese PLA, Chengdu, China and plasmids by Dr. Ma, Ocean University of China, Qingdao, China.

Conflict of interest


Supplementary material

11060_2012_1033_MOESM1_ESM.ppt (626 kb)
Supplementary material 1 (PPT 626 kb)
11060_2012_1033_MOESM2_ESM.docx (11 kb)
Supplementary material 2 (DOCX 11 kb)


  1. 1.
    Reardon DA, Galanis E, DeGroot JF, Cloughesy TF, Wefel JS, Lamborn KR, Lassman AB, Gilbert MR, Sampson JH, Wick W et al (2011) Clinical trial end points for high-grade glioma: the evolving landscape. Neuro Oncol 13(3):353–361Google Scholar
  2. 2.
    Vecil GG, Lang FF (2003) Clinical trials of adenoviruses in brain tumors: a review of Ad-p53 and oncolytic adenoviruses. J Neurooncol 65(3):237–246PubMedCrossRefGoogle Scholar
  3. 3.
    Shewach DS, Zerbe LK, Hughes TL, Roessler BJ, Breakefield XO, Davidson BL (1994) Enhanced cytotoxicity of antiviral drugs mediated by adenovirus directed transfer of the herpes simplex virus thymidine kinase gene in rat glioma cells. Cancer Gene Ther 1(2):107–112PubMedGoogle Scholar
  4. 4.
    McBride WH (2012) Integration of adenovirus thymidine kinase suicide-gene therapy with surgery and radiation therapy for malignant glioma. Future Oncol 8(1):17–20Google Scholar
  5. 5.
    Fueyo J, Gomez-Manzano C, Yung WK, Clayman GL, Liu TJ, Bruner J, Levin VA, Kyritsis AP (1996) Adenovirus-mediated p16/CDKN2 gene transfer induces growth arrest and modifies the transformed phenotype of glioma cells. Oncogene 12(1):103–110PubMedGoogle Scholar
  6. 6.
    Chintala SK, Fueyo J, Gomez-Manzano C, Venkaiah B, Bjerkvig R, Yung WK, Sawaya R, Kyritsis AP, Rao JS (1997) Adenovirus-mediated p16/CDKN2 gene transfer suppresses glioma invasion in vitro. Oncogene 15(17):2049–2057PubMedCrossRefGoogle Scholar
  7. 7.
    Fueyo J, Gomez-Manzano C, Puduvalli VK, Martin-Duque P, Perez-Soler R, Levin VA, Yung WK, Kyritsis AP (1998) Adenovirus-mediated p16 transfer to glioma cells induces G1 arrest and protects from paclitaxel and topotecan: implications for therapy. Int J Oncol 12(3):665–669PubMedGoogle Scholar
  8. 8.
    Lee SH, Kim MS, Kwon HC, Park IC, Park MJ, Lee CT, Kim YW, Kim CM, Hong SI (2000) Growth inhibitory effect on glioma cells of adenovirus-mediated p16/INK4a gene transfer in vitro and in vivo. Int J Mol Med 6(5):559–563PubMedGoogle Scholar
  9. 9.
    Liu J, Xu X, Feng X, Zhang B, Wang J (2011) Adenovirus-mediated delivery of bFGF small interfering RNA reduces STAT3 phosphorylation and induces the depolarization of mitochondria and apoptosis in glioma cells U251. J Exp Clin Cancer Res 30:80Google Scholar
  10. 10.
    Zhang J, Zhang QY, Fu YC, Wang T, Zhang J, Xu P, Zhou X, Pu PY, Kang CS (2009) Expression of p-Akt and COX-2 in gastric adenocarcinomas and adenovirus mediated Akt1 and COX-2 ShRNA suppresses SGC-7901 gastric adenocarcinoma and U251 glioma cell growth in vitro and in vivo. Technol Cancer Res Treat 8(6):467–478PubMedGoogle Scholar
  11. 11.
    Pan D, Wei X, Liu M, Feng S, Tian X, Feng X, Zhang X (2010) Adenovirus mediated transfer of p53, GM-CSF and B7-1 suppresses growth and enhances immunogenicity of glioma cells. Neurol Res 32(5):502–509Google Scholar
  12. 12.
    Naumann U, Bahr O, Wolburg H, Altenberend S, Wick W, Liston P, Ashkenazi A, Weller M (2007) Adenoviral expression of XIAP antisense RNA induces apoptosis in glioma cells and suppresses the growth of xenografts in nude mice. Gene Ther 14(2):147–161PubMedCrossRefGoogle Scholar
  13. 13.
    Lee J, Hampl M, Albert P, Fine HA (2002) Antitumor activity and prolonged expression from a TRAIL-expressing adenoviral vector. Neoplasia 4(4):312–323PubMedCrossRefGoogle Scholar
  14. 14.
    Kim CY, Jeong M, Mushiake H, Kim BM, Kim WB, Ko JP, Kim MH, Kim M, Kim TH, Robbins PD et al (2006) Cancer gene therapy using a novel secretable trimeric TRAIL. Gene Ther 13(4):330–338PubMedCrossRefGoogle Scholar
  15. 15.
    Wohlfahrt ME, Beard BC, Lieber A, Kiem HP (2007) A capsid-modified, conditionally replicating oncolytic adenovirus vector expressing TRAIL Leads to enhanced cancer cell killing in human glioblastoma models. Cancer Res 67(18):8783–8790PubMedCrossRefGoogle Scholar
  16. 16.
    Jeong M, Kwon YS, Park SH, Kim CY, Jeun SS, Song KW, Ko Y, Robbins PD, Billiar TR, Kim BM et al (2009) Possible novel therapy for malignant gliomas with secretable trimeric TRAIL. PLoS One 4(2):e4545PubMedCrossRefGoogle Scholar
  17. 17.
    Liu Y, Lang F, Xie X, Prabhu S, Xu J, Sampath D, Aldape K, Fuller G, Puduvalli VK (2011) Efficacy of adenovirally expressed soluble TRAIL in human glioma organotypic slice culture and glioma xenografts. Cell Death Dis 2:e121Google Scholar
  18. 18.
    Li JT, Bian K, Zhang AL, Kim DH, Ashley WW, Nath R, McCutcheon I, Fang B, Murad F (2011) Targeting different types of human meningioma and glioma cells using a novel adenoviral vector expressing GFP-TRAIL fusion protein from hTERT promoter. Cancer Cell Int 11(1):35Google Scholar
  19. 19.
    Finnberg N, El-Deiry WS (2008) TRAIL death receptors as tumor suppressors and drug targets. Cell Cycle 7(11):1525–1528PubMedCrossRefGoogle Scholar
  20. 20.
    Sontheimer EJ, Carthew RW (2005) Silence from within: endogenous siRNAs and miRNAs. Cell 122(1):9–12PubMedCrossRefGoogle Scholar
  21. 21.
    Lovat F, Valeri N, Croce CM (2011) MicroRNAs in the pathogenesis of cancer. Semin Oncol 38(6):724–733Google Scholar
  22. 22.
    Varol N, Konac E, Gurocak OS, Sozen S (2011) The realm of microRNAs in cancers. Mol Biol Rep 38(2):1079–1089Google Scholar
  23. 23.
    Zhang G, Wang Q, Xu R (2011) Therapeutics based on microRNA: a new approach for liver cancer. Curr Genomics 11(5):311–325Google Scholar
  24. 24.
    Skalsky RL, Cullen BR (2011) Reduced expression of brain-enriched microRNAs in glioblastomas permits targeted regulation of a cell death gene. PLoS One 6(9):e24248Google Scholar
  25. 25.
    Silber J, Lim DA, Petritsch C, Persson AI, Maunakea AK, Yu M, Vandenberg SR, Ginzinger DG, James CD, Costello JF et al (2008) miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells. BMC Med 6:14Google Scholar
  26. 26.
    Li KK, Pang JC, Ching AK, Wong CK, Kong X, Wang Y, Zhou L, Chen Z, Ng HK (2009) miR-124 is frequently down-regulated in medulloblastoma and is a negative regulator of SLC16A1. Hum Pathol 40(9):1234–1243PubMedCrossRefGoogle Scholar
  27. 27.
    Li D, Chen P, Li XY, Zhang LY, Xiong W, Zhou M, Xiao L, Zeng F, Li XL, Wu MH et al (2011) Grade-specific expression profiles of miRNAs/mRNAs and docking study in human grade I-III astrocytomas. Omics 15(10):673–682Google Scholar
  28. 28.
    Xia H, Cheung WK, Ng SS, Jiang X, Jiang S, Sze J, Leung GK, Lu G, Chan DT, Bian XW et al (2012) Loss of brain-enriched miR-124 MicroRNA enhances stem-like traits and invasiveness of glioma cells. J Biol Chem 287(13):9962–9971Google Scholar
  29. 29.
    Papagiannakopoulos T, Friedmann-Morvinski D, Neveu P, Dugas JC, Gill RM, Huillard E, Liu C, Zong H, Rowitch DH, Barres BA et al (2012) Pro-neural miR-128 is a glioma tumor suppressor that targets mitogenic kinases. Oncogene 31(15):1884–1895Google Scholar
  30. 30.
    Xia H, Qi Y, Ng SS, Chen X, Li D, Chen S, Ge R, Jiang S, Li G, Chen Y et al (2009) microRNA-146b inhibits glioma cell migration and invasion by targeting MMPs. Brain Res 1269:158–165PubMedCrossRefGoogle Scholar
  31. 31.
    Yue X, Wang P, Xu J, Zhu Y, Sun G, Pang Q, Tao R (2012) MicroRNA-205 functions as a tumor suppressor in human glioblastoma cells by targeting VEGF-A. Oncol Rep 27(4):1200–1206Google Scholar
  32. 32.
    Rao SA, Santosh V, Somasundaram K (2010) Genome-wide expression profiling identifies deregulated miRNAs in malignant astrocytoma. Mod Pathol 23(10):1404–1417Google Scholar
  33. 33.
    Song L, Huang Q, Chen K, Liu L, Lin C, Dai T, Yu C, Wu Z, Li J (2010) miR-218 inhibits the invasive ability of glioma cells by direct downregulation of IKK-beta. Biochem Biophys Res Commun 402(1):135–140Google Scholar
  34. 34.
    Liu Y, Yan W, Zhang W, Chen L, You G, Bao Z, Wang Y, Wang H, Kang C, Jiang T (2012) MiR-218 reverses high invasiveness of glioblastoma cells by targeting the oncogenic transcription factor LEF1. Oncol Rep 28(3):1013–1021Google Scholar
  35. 35.
    Katakowski M, Zheng X, Jiang F, Rogers T, Szalad A, Chopp M (2010) MiR-146b-5p suppresses EGFR expression and reduces in vitro migration and invasion of glioma. Cancer Invest 28(10):1024–1030Google Scholar
  36. 36.
    Pierson J, Hostager B, Fan R, Vibhakar R (2008) Regulation of cyclin dependent kinase 6 by microRNA 124 in medulloblastoma. J Neurooncol 90(1):1–7PubMedCrossRefGoogle Scholar
  37. 37.
    Zhang Y, Chao T, Li R, Liu W, Chen Y, Yan X, Gong Y, Yin B, Liu W, Qiang B et al (2009) MicroRNA-128 inhibits glioma cells proliferation by targeting transcription factor E2F3a. J Mol Med (Berl) 87(1):43–51CrossRefGoogle Scholar
  38. 38.
    Godlewski J, Nowicki MO, Bronisz A, Williams S, Otsuki A, Nuovo G, Raychaudhury A, Newton HB, Chiocca EA, Lawler S (2008) Targeting of the Bmi-1 oncogene/stem cell renewal factor by microRNA-128 inhibits glioma proliferation and self-renewal. Cancer Res 68(22):9125–9130PubMedCrossRefGoogle Scholar
  39. 39.
    McCutcheon IE, Friend KE, Gerdes TM, Zhang BM, Wildrick DM, Fuller GN (2000) Intracranial injection of human meningioma cells in athymic mice: an orthotopic model for meningioma growth. J Neurosurg 92(2):306–314PubMedCrossRefGoogle Scholar
  40. 40.
    Klapper W, Shin T, Mattson MP (2001) Differential regulation of telomerase activity and TERT expression during brain development in mice. J Neurosci Res 64(3):252–260PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Department of NeurosurgeryThe Affiliated Hospital of Medical College, Qingdao UniversityQingdaoChina

Personalised recommendations