, Volume 28, Issue 1, pp 41–54 | Cite as

miR-125b Inhibitor May Enhance the Invasion-Prevention Activity of Temozolomide in Glioblastoma Stem Cells by Targeting PIAS3

  • Lei Shi
  • Yi Wan
  • Guan Sun
  • Shuguang Zhang
  • Zhimin WangEmail author
  • Yanjun ZengEmail author
Original Research Article



Temozolomide, an alkylating agent, is a promising chemotherapeutic agent for treating glioblastoma. Although chemotherapy with temozolomide may restrain tumor growth for some months, invariable tumor recurrence suggests that cancer stem cells maintaining these tumors persist. Previous research has shown that temozolomide can inhibit the proliferation of human glioblastoma stem cells (GSCs); however, no research has focused on the invasion of GSCs, which is an important factor for glioblastoma recurrence. Accumulating evidence indicates that microRNA (miR)-125b over-expression in GSCs may increase their invasiveness.


Our objective was to identify the effects and mechanism of action of an miR-125b inhibitor combined with temozolomide in the invasive pathogenesis of GSCs.


We modified the levels of miR-125b expression in primary GSCs in order to observe the effect on sensitivity to temozolomide on invasion, and we further analyzed the differences in mechanism between miR-125b treatment alone and treatment with miR-125b plus temozolomide using the Cancer PathwayFinder PCR Array.


Our results demonstrated that either an miR-125b inhibitor or temozolomide could modestly inhibit the invasiveness of GSCs. Furthermore, GSCs that were pre-transfected with an miR-125b inhibitor, then treated with temozolomide, showed significantly decreased invasiveness when compared with GSCs treated with an miR-125b inhibitor or temozolomide alone. Further research into the underlying mechanism demonstrated that the miR-125b inhibitor enhanced the invasion-prevention activity of temozolomide in GSCs through targeting PIAS3 (protein inhibitor of activated STAT [signal transducer and activator of transcription]), which contributed to reduced STAT3 transcriptional activity and subsequent decreased expression of matrix metalloproteinase (MMP)-2 and -9.


miR-125b could play a role in the development of temozolomide resistance in GSCs. Inhibition of miR-125b expression may enhance sensitivity of GSCs to temozolomide by targeting PIAS3 on cell invasion.


Reverse Transcription Polymerase Chain Reaction Temozolomide Temozolomide Treatment Glioblastoma Stem Cell Chemotherapeutic Sensitivity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported by the China Natural Science Foundation (81000963 and 81201976), Jiangsu Province’s 333 Talent Program (BRA2011046), Jiangsu Province’s Natural Science Foundation (BK2012670), Medical Research Foundation by Jiangsu Province Health Department (YG201301 and Z201318), the Clinical Technology Development of Jiangsu University (JLY20120053), the Kunshan Social Development Foundation (KS1006, KS1009), and the Suzhou Social Development Foundation (SYS201063).

Conflicts of interest

All authors have declared the sources of research funding for this manuscript and have no financial or other contractual agreements that might cause (or be perceived as causes of) conflicts of interest.


  1. 1.
    Stupp R, Tonn JC, Brada M, et al. High-grade malignant glioma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2010;21:v190–3.PubMedCrossRefGoogle Scholar
  2. 2.
    Clarke MJ, Mulligan EA, Grogan PT, et al. Effective sensitization of temozolomide by ABT-888 is lost with development of temozolomide resistance in glioblastoma xenograft lines. Mol Cancer Ther. 2009;8:407–14.PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Athanassiou H, Synodinou M, Maragoudakis E, et al. Randomized phase II study of temozolomide and radiotherapy compared with radiotherapy alone in newly diagnosed glioblastoma multiforme. J Clin Oncol. 2005;23:2372–7.PubMedCrossRefGoogle Scholar
  4. 4.
    Mei J, Bachoo R, Zhang C. MicroRNA-146a inhibits glioma development by targeting Notch1. Mol Cell Biol. 2011;31:3584–92.PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Hatfield SD, Shcherbata HR, Fischer KA, et al. Stem cell division is regulated by the microRNA pathway. Nature. 2005;435:974–8.PubMedCrossRefGoogle Scholar
  6. 6.
    Shi L, Zhang J, Pan T, et al. MiR-125b is critical for the suppression of human U251 glioma stem cell proliferation. Brain Res. 2010;1312:120–6.PubMedCrossRefGoogle Scholar
  7. 7.
    Mishra PJ, Bertino JR. MicroRNA polymorphisms: the future of pharmacogenomics, molecular epidemiology and individualized medicine. Pharmacogenomics. 2009;10:399–416.PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Rukov JL, Vinther J, Shomron N. Pharmacogenomics genes show varying perceptibility to microRNA regulation. Pharmacogenet Genomics. 2011;21:251–62.PubMedCrossRefGoogle Scholar
  9. 9.
    Shomron N. MicroRNAs and pharmacogenomics. Pharmacogenomics. 2010;11:629–32.PubMedCrossRefGoogle Scholar
  10. 10.
    Ren Y, Zhou X, Mei M, et al. MicroRNA-21 inhibitor sensitizes human glioblastoma cells U251 (PTEN-mutant) and LN229 (PTEN-wild type) to taxol. BMC Cancer. 2010;10:27–39.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Shi L, Zhang S, Feng K, et al. MicroRNA-125b-2 confers human glioblastoma stem cells resistance to temozolomide through the mitochondrial pathway of apoptosis. Int J Oncol. 2012;40(1):119–29.PubMedGoogle Scholar
  12. 12.
    Zhang S, Wan Y, Pan T, et al. MicroRNA-21 inhibitor sensitizes human glioblastoma U251 stem cells to chemotherapeutic drug temozolomide. J Mol Neurosci. 2012;47(2):346–56.PubMedCrossRefGoogle Scholar
  13. 13.
    Darling JL. The in vitro biology of human brain tumors. In: Thomas DGT, editor. Neuro-oncology: primary malignant brain tumors. Baltimore (MD): Johns Hopkins University Press; 1990: 1–25.Google Scholar
  14. 14.
    Chen C, Ridzon DA, Broomer AJ, et al. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005;33:e179.PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (−Delta Delta C(T)) Method. Methods. 2001;25:402–8.PubMedCrossRefGoogle Scholar
  16. 16.
    Shi L, Wan Y, Sun G, et al. Functional differences of miR-125b on the invasion of primary glioblastoma CD133-negative cells and CD133-positive cells. Neuromol Med. 2012;14(4):303–16.CrossRefGoogle Scholar
  17. 17.
    Xia HF, He TZ, Liu CM, et al. MiR-125b expression affects the proliferation and apoptosis of human glioma cells by targeting Bmf. Cell Physiol Biochem. 2009;23:347–58.PubMedCrossRefGoogle Scholar
  18. 18.
    Brantley EC, Nabors LB, Gillespie GY, et al. Loss of protein inhibitors of activated STAT-3 expression in glioblastoma multiforme tumors: implications for STAT-3 activation and gene expression. Clin Cancer Res. 2008;14:4694–704.PubMedCrossRefGoogle Scholar
  19. 19.
    Wei D, Le X, Zheng L, et al. Stat3 activation regulates the expression of vascular endothelial growth factor and human pancreatic cancer angiogenesis and metastasis. Oncogene. 2003;22(3):319–29.PubMedCrossRefGoogle Scholar
  20. 20.
    Pan TH, Wang YY, Zhang SG, et al. Research on the effects of PIAS3 expression on the invasion of glioma TJ905 cells. Zhonghua Wai Ke Za Zhi. 2011;49(5):440–4.PubMedGoogle Scholar
  21. 21.
    Johnson CD, Esquela-Kerscher A, Stefani G, et al. The let-7 microrna represses cell proliferation pathways in human cells. Cancer Res. 2007;67:7713–22.PubMedCrossRefGoogle Scholar
  22. 22.
    Wan Y, Fei XF, Wang ZM, et al. Expression of miR-125b in the new, highly invasive glioma stem cell and progenitor cell line SU3. Chin J Cancer. 2012;31(4):207–14.PubMedCrossRefGoogle Scholar
  23. 23.
    Cairo S, Wang Y, de Reyniès A, et al. Stem cell-like micro-RNA signature driven by Myc in aggressive liver cancer. PNAS. 2010;107:20471–6.PubMedCrossRefGoogle Scholar
  24. 24.
    Vescovi AL, Galli R, Reynolds BA. Brain tumour stem cells. Nat Rev Cancer. 2006;6:425–36.PubMedCrossRefGoogle Scholar
  25. 25.
    Wang L, Zhang J, Banerjee S, et al. Sumoylation of vimentin354 is associated with PIAS3 inhibition of glioma cell migration. Oncotarget. 2010;1(7):620–7.PubMedGoogle Scholar
  26. 26.
    Zhang X, Sun Y, Pireddu R, et al. A novel inhibitor of STAT3 homodimerization selectively suppresses STAT3 activity and malignant transformation. Cancer Res. 2013;73(6):1922–33.PubMedCrossRefGoogle Scholar
  27. 27.
    Senft C, Priester M, Polacin M, et al. Inhibition of the JAK-2/STAT3 signaling pathway impedes the migratory and invasive potential of human glioblastoma cells. J Neurooncol. 2011;101(3):393–403.PubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2013

Authors and Affiliations

  1. 1.Department of NeurosurgeryThe First People’s Hospital of Kunshan affiliated with Jiangsu UniversitySuzhouPeople’s Republic of China
  2. 2.Department of NeurosurgerySuzhou Kowloon Hospital affiliated with Shanghai Jiao Tong University School of MedicineSuzhouPeople’s Republic of China
  3. 3.Department of NeurosurgeryFourth Affiliated Yancheng Hospital of Nantong UniversityYanchengPeople’s Republic of China
  4. 4.Beijing University of TechnologyBeijingPeople’s Republic of China

Personalised recommendations