Molecular and Cellular Biochemistry

, Volume 403, Issue 1–2, pp 33–41 | Cite as

MiR-1224-5p acts as a tumor suppressor by targeting CREB1 in malignant gliomas

  • Jin Qian
  • Rui Li
  • Ying-Yi Wang
  • Yan Shi
  • Wen-Kang Luan
  • Tao Tao
  • Jun-Xia Zhang
  • Yi-Chang Xu
  • Yong-Ping You


The dysregulation of miR-1224-5p has been reported in several human cancers. However, the expression and function of miR-1224-5p in glioma remains unknown. The aim of our study was to investigate the effect of miR-1224-5p on glioma cells and to determine its functional signaling mediators. Using 198 glioma samples within the Chinese Glioma Genome Atlas expression dataset, we demonstrated that miR-1224-5p expression is decreased in high-grade gliomas when compared with low-grade gliomas. Differential miR-1224-5p expression in 50 randomly selected samples was verified by in situ hybridization. The expression of miR-1224-5p was shown to positively correlate with overall survival in 82 glioblastoma patients. Exogenous expression of miR-1224-5p in glioma cells suppressed proliferation and invasion and promoted apoptosis. Target prediction algorithms identified a consensus miR-1224-5p recognition site in the 3′UTR of the cAMP response element-binding protein (CREB1) gene, and this sequence was shown to directly confer miR-1224-5p repression in luciferase reporter assays. Furthermore, exogenous miR-1224-5p expression was shown to down-regulate CREB1, as well as its downstream target genes matrix metalloproteinase-9 and B-cell lymphoma-2. Conversely, over-expression of CREB1 reversed the effect of miR-1224-5p on the proliferation, invasion, and apoptosis of glioma cells. These data indicate that miR-1224-5p may inhibit tumor-associated activity in malignant gliomas by targeting CREB1.


MiR-1224-5p CREB1 Glioma Cell proliferation Invasion Apoptosis 



This work was supported by grants from the National High Technology Research and Development Program of China (863) (2012AA02A508), the Research Special Fund For Public Welfare Industry of Health (201402008), National Natural Science Foundation of China (91229121, 81272792, 81472362, 81172389, 81372709, 81302185), Jiangsu Province’s Natural Science Foundation (20131019), Jiangsu Province’s Key Provincial Talents Program (RC2011051), Jiangsu Province’s Key Discipline of Medicine (XK201117), Jiangsu Provincial Special Program of Medical Science (BL2012028), and Program for Development of Innovative Research Team in the First Affiliated Hospital of NJMU, and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Supplementary material

11010_2015_2334_MOESM1_ESM.docx (10 kb)
Supplementary material 1 (DOCX 10 kb)
11010_2015_2334_MOESM2_ESM.tif (73 kb)
Supplementary material 2 (TIFF 73 kb)


  1. 1.
    Clarke J, Butowski N, Chang S (2010) Recent advances in therapy for glioblastoma. Arch Neurol 67:279–283. doi: 10.1001/archneurol.2010.5 CrossRefPubMedGoogle Scholar
  2. 2.
    Saika K, Katanoda K (2011) Comparison of time trends in brain and central nervous system cancer mortality (1990–2006) between countries based on the WHO mortality database. Jpn J Clin Oncol 41:304–305. doi: 10.1093/jjco/hyr004 CrossRefPubMedGoogle Scholar
  3. 3.
    Yan W, Zhang W, Jiang T (2011) Oncogene addiction in gliomas: implications for molecular targeted therapy. J Exp Clin Cancer Res 30:58. doi: 10.1186/1756-9966-30-58 CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Tate MC, Aghi MK (2009) Biology of angiogenesis and invasion in glioma. Neurotherapeutics 6:447–457. doi: 10.1016/j.nurt.2009.04.001 CrossRefPubMedGoogle Scholar
  5. 5.
    Yue J, Tigyi G (2006) MicroRNA trafficking and human cancer. Cancer Biol Ther 5:573–578CrossRefPubMedGoogle Scholar
  6. 6.
    Zimmerman AL, Wu S (2011) MicroRNAs, cancer and cancer stem cells. Cancer Lett 300:10–19. doi: 10.1016/j.canlet.2010.09.019 CrossRefPubMedCentralPubMedGoogle Scholar
  7. 7.
    Zhang CM, Zhao J, Deng HY (2013) MiR-155 promotes proliferation of human breast cancer MCF-7 cells through targeting tumor protein 53-induced nuclear protein 1. J Biomed Sci 20:79. doi: 10.1186/1423-0127-20-79 CrossRefPubMedCentralPubMedGoogle Scholar
  8. 8.
    Wang XF, Shi ZM, Wang XR, Cao L, Wang YY, Zhang JX, Yin Y, Luo H, Kang CS, Liu N, Jiang T, You YP (2012) MiR-181d acts as a tumor suppressor in glioma by targeting K-ras and Bcl-2. J Cancer Res Clin Oncol 138:573–584. doi: 10.1007/s00432-011-1114-x CrossRefPubMedGoogle Scholar
  9. 9.
    Zhang C, Zhang J, Hao J, Shi Z, Wang Y, Han L, Yu S, You Y, Jiang T, Wang J, Liu M, Pu P, Kang C (2012) High level of miR-221/222 confers increased cell invasion and poor prognosis in glioma. J Transl Med 10:119. doi: 10.1186/1479-5876-10-119 CrossRefPubMedCentralPubMedGoogle Scholar
  10. 10.
    Krishnan K, Steptoe AL, Martin HC, Pattabiraman DR, Nones K, Waddell N, Mariasegaram M, Simpson PT, Lakhani SR, Vlassov A, Grimmond SM, Cloonan N (2013) MiR-139-5p is a regulator of metastatic pathways in breast cancer. RNA. doi: 10.1261/rna.042143.113 Google Scholar
  11. 11.
    Spaccarotella E, Pellegrino E, Ferracin M, Ferreri C, Cuccuru G, Liu C, Iqbal J, Cantarella D, Taulli R, Provero P, Di Cunto F, Medico E, Negrini M, Chan WC, Inghirami G, Piva R (2013) STAT3-mediated activation of microRNA cluster 17–92 promotes proliferation and survival of ALK positive anaplastic large cell lymphoma. Haematologica. doi: 10.3324/haematol.2013.088286 PubMedGoogle Scholar
  12. 12.
    Tokarz P, Blasiak J (2012) The role of microRNA in metastatic colorectal cancer and its significance in cancer prognosis and treatment. Acta Biochim Pol 59:467–474PubMedGoogle Scholar
  13. 13.
    Garofalo M, Jeon YJ, Nuovo GJ, Middleton J, Secchiero P, Joshi P, Alder H, Nazaryan N, Di Leva G, Romano G, Crawford M, Nana-Sinkam P, Croce CM (2013) MiR-34a/c-dependent PDGFR-alpha/beta downregulation inhibits tumorigenesis and enhances TRAIL-induced apoptosis in lung cancer. PLoS One 8:e67581. doi: 10.1371/journal.pone.0067581 CrossRefPubMedCentralPubMedGoogle Scholar
  14. 14.
    Feliciano A, Castellvi J, Artero-Castro A, Leal JA, Romagosa C, Hernandez-Losa J, Peg V, Fabra A, Vidal F, Kondoh H, Ramon YCS, Lleonart ME (2013) MiR-125b acts as a tumor suppressor in breast tumorigenesis via Its novel direct targets ENPEP, CK2-alpha, CCNJ, and MEGF9. PLoS One 8:e76247. doi: 10.1371/journal.pone.0076247 CrossRefPubMedCentralPubMedGoogle Scholar
  15. 15.
    Kishikawa T, Otsuka M, Yoshikawa T, Ohno M, Takata A, Shibata C, Kondo Y, Akanuma M, Yoshida H, Koike K (2013) Regulation of the expression of the liver cancer susceptibility gene MICA by microRNAs. Sci Rep 3:2739. doi: 10.1038/srep02739 CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    Rao SA, Arimappamagan A, Pandey P, Santosh V, Hegde AS, Chandramouli BA, Somasundaram K (2013) MiR-219-5p inhibits receptor tyrosine kinase pathway by targeting EGFR in glioblastoma. PLoS One 8:e63164. doi: 10.1371/journal.pone.0063164 CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Mosakhani N, Lahti L, Borze I, Karjalainen-Lindsberg ML, Sundstrom J, Ristamaki R, Osterlund P, Knuutila S, Sarhadi VK (2012) MicroRNA profiling predicts survival in anti-EGFR treated chemorefractory metastatic colorectal cancer patients with wild-type KRAS and BRAF. Cancer Genet 205:545–551. doi: 10.1016/j.cancergen.2012.08.003 CrossRefPubMedGoogle Scholar
  18. 18.
    Della Vittoria Scarpati G, Falcetta F, Carlomagno C, Ubezio P, Marchini S, De Stefano A, Singh VK, D’Incalci M, De Placido S, Pepe S (2012) A specific miRNA signature correlates with complete pathological response to neoadjuvant chemoradiotherapy in locally advanced rectal cancer. Int J Radiat Oncol Biol Phys 83:1113–1119. doi: 10.1016/j.ijrobp.2011.09.030 CrossRefPubMedGoogle Scholar
  19. 19.
    Nymark P, Guled M, Borze I, Faisal A, Lahti L, Salmenkivi K, Kettunen E, Anttila S, Knuutila S (2011) Integrative analysis of microRNA, mRNA and aCGH data reveals asbestos- and histology-related changes in lung cancer. Genes Chromosom Cancer 50:585–597. doi: 10.1002/gcc.20880 CrossRefPubMedGoogle Scholar
  20. 20.
    Park JK, Park SH, So K, Bae IH, Yoo YD, Um HD (2010) ICAM-3 enhances the migratory and invasive potential of human non-small cell lung cancer cells by inducing MMP-2 and MMP-9 via Akt and CREB. Int J Oncol 36:181–192PubMedGoogle Scholar
  21. 21.
    Zhang M, Xu JJ, Zhou RL, Zhang QY (2013) cAMP responsive element binding protein-1 is a transcription factor of lysosomal-associated protein transmembrane-4 beta in human breast cancer cells. PLoS One 8:e57520. doi: 10.1371/journal.pone.0057520 CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Pigazzi M, Manara E, Baron E, Basso G (2009) miR-34b targets cyclic AMP-responsive element binding protein in acute myeloid leukemia. Cancer Res 69:2471–2478. doi: 10.1158/0008-5472.CAN-08-3404 CrossRefPubMedGoogle Scholar
  23. 23.
    Perry C, Sklan EH, Soreq H (2004) CREB regulates AChE-R-induced proliferation of human glioblastoma cells. Neoplasia 6:279–286. doi: 10.1593/neo.3424 CrossRefPubMedCentralPubMedGoogle Scholar
  24. 24.
    Sakamoto KM, Frank DA (2009) CREB in the pathophysiology of cancer: implications for targeting transcription factors for cancer therapy. Clin Cancer Res 15:2583–2587. doi: 10.1158/1078-0432.CCR-08-1137 CrossRefPubMedCentralPubMedGoogle Scholar
  25. 25.
    Shukla A, Bosenberg MW, MacPherson MB, Butnor KJ, Heintz NH, Pass HI, Carbone M, Testa JR, Mossman BT (2009) Activated cAMP response element binding protein is overexpressed in human mesotheliomas and inhibits apoptosis. Am J Pathol 175:2197–2206. doi: 10.2353/ajpath.2009.090400 CrossRefPubMedCentralPubMedGoogle Scholar
  26. 26.
    Peng B, Hu S, Jun Q, Luo D, Zhang X, Zhao H, Li D (2013) MicroRNA-200b targets CREB1 and suppresses cell growth in human malignant glioma. Mol Cell Biochem 379:51–58. doi: 10.1007/s11010-013-1626-6 CrossRefPubMedGoogle Scholar
  27. 27.
    Van Themsche C, Mathieu I, Parent S, Asselin E (2007) Transforming growth factor-beta3 increases the invasiveness of endometrial carcinoma cells through phosphatidylinositol 3-kinase-dependent up-regulation of X-linked inhibitor of apoptosis and protein kinase c-dependent induction of matrix metalloproteinase-9. J Biol Chem 282:4794–4802. doi: 10.1074/jbc.M608497200 CrossRefPubMedGoogle Scholar
  28. 28.
    Liu G, Ding W, Neiman J, Mulder KM (2006) Requirement of Smad3 and CREB-1 in mediating transforming growth factor-beta (TGF beta) induction of TGF beta 3 secretion. J Biol Chem 281:29479–29490. doi: 10.1074/jbc.M600579200 CrossRefPubMedGoogle Scholar
  29. 29.
    Perianayagam MC, Madias NE, Pereira BJ, Jaber BL (2006) CREB transcription factor modulates Bcl2 transcription in response to C5a in HL-60-derived neutrophils. Eur J Clin Invest 36:353–361. doi: 10.1111/j.1365-2362.2006.01637.x CrossRefPubMedGoogle Scholar
  30. 30.
    Mendell JT (2005) MicroRNAs: critical regulators of development, cellular physiology and malignancy. Cell Cycle 4:1179–1184CrossRefPubMedGoogle Scholar
  31. 31.
    Hwang HW, Mendell JT (2007) MicroRNAs in cell proliferation, cell death, and tumorigenesis. Br J Cancer 96(Suppl):R40–R44PubMedGoogle Scholar
  32. 32.
    Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, Sweet-Cordero A, Ebert BL, Mak RH, Ferrando AA, Downing JR, Jacks T, Horvitz HR, Golub TR (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838. doi: 10.1038/nature03702 CrossRefPubMedGoogle Scholar
  33. 33.
    Xi JJ (2013) MicroRNAs in cancer. Cancer Treat Res 158:119–137. doi: 10.1007/978-3-642-31659-3_5 CrossRefPubMedGoogle Scholar
  34. 34.
    Mayr B, Montminy M (2001) Transcriptional regulation by the phosphorylation-dependent factor CREB. Nat Rev Mol Cell Biol 2:599–609. doi: 10.1038/35085068 CrossRefPubMedGoogle Scholar
  35. 35.
    Shaywitz AJ, Greenberg ME (1999) CREB: a stimulus-induced transcription factor activated by a diverse array of extracellular signals. Annu Rev Biochem 68:821–861. doi: 10.1146/annurev.biochem.68.1.821 CrossRefPubMedGoogle Scholar
  36. 36.
    Majumder S, Varadharaj S, Ghoshal K, Monani U, Burghes AH, Jacob ST (2004) Identification of a novel cyclic AMP-response element (CRE-II) and the role of CREB-1 in the cAMP-induced expression of the survival motor neuron (SMN) gene. J Biol Chem 279:14803–14811. doi: 10.1074/jbc.M308225200 CrossRefPubMedCentralPubMedGoogle Scholar
  37. 37.
    Alvarez-Lopez C, Cernuda-Cernuda R, Paniagua MA, Alvarez-Viejo M, Fernandez-Lopez A, Garcia-Fernandez JM (2004) The transcription factor CREB is phosphorylated in neurons of the piriform cortex of blind mice in response to illumination of the retina. Neurosci Lett 357:223–226. doi: 10.1016/j.neulet.2003.12.099 CrossRefPubMedGoogle Scholar
  38. 38.
    Thway K, Fisher C (2012) Tumors with EWSR1-CREB1 and EWSR1-ATF1 fusions: the current status. Am J Surg Pathol 36:e1–e11. doi: 10.1097/PAS.0b013e31825485c5 CrossRefPubMedGoogle Scholar
  39. 39.
    Tan X, Wang S, Zhu L, Wu C, Yin B, Zhao J, Yuan J, Qiang B, Peng X (2012) cAMP response element-binding protein promotes gliomagenesis by modulating the expression of oncogenic microRNA-23a. Proc Natl Acad Sci USA 109:15805–15810. doi: 10.1073/pnas.1207787109 CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Jin Qian
    • 1
    • 2
  • Rui Li
    • 1
  • Ying-Yi Wang
    • 1
  • Yan Shi
    • 1
    • 3
  • Wen-Kang Luan
    • 1
  • Tao Tao
    • 1
  • Jun-Xia Zhang
    • 1
  • Yi-Chang Xu
    • 2
  • Yong-Ping You
    • 1
  1. 1.Department of NeurosurgeryThe First Affiliated Hospital of Nanjing Medical UniversityNanjingChina
  2. 2.Department of NeurosurgeryThe People’s Hospital of Xuancheng CityXuanchengChina
  3. 3.Department of Neurosurgery, Nanjing First HospitalNanjing Medical UniversityNanjingChina

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