Abstract
Glioblastoma multiforme (GBM), the most common brain tumor in adults, is neurologically destructive and has a dismal response to virtually all therapeutic modalities. One phenomenon that can contribute to this complexity is the presence of a relatively small subset of glioma stem cells (GSCs) within the tumor and the activation of pathways that control cellular differentiation. The Notch signaling pathway, which is responsible for maintaining a balance between cell proliferation and apoptosis, is believed to be deregulated in cancer stem cells (CSCs), leading to tumor growth through the generation or expansion of CSCs. In this study, Notch-1 small interfering RNA (siRNA) was used to silence Notch-1 gene expression in GSCs. An MTT assay demonstrated inhibitory effects on the proliferation of GSCs in vitro. Real-time PCR showed that Notch-1 expression levels were markedly decreased in GSCs transfected with Notch-1 siRNA in vitro. Notch-1 silenced GSCs engrafted on Balb/c nude mice showed a significantly greater reduction in oncogenicity than the control group (P < 0.05). Furthermore, direct intratumoral injections of Notch-1-siRNA/PEI significantly delayed the growth of pre-established tumors in nude mice (P < 0.05). These results suggest that siRNA-mediated silencing of the Notch-1 gene may represent a novel target for gene therapy of GBM.
Similar content being viewed by others
References
Porter KR, McCarthy BJ, Freels S et al (2010) Prevalence estimates for primary brain tumors in the United States by age, gender, behavior, and histology. Neuro Oncol 12:520–527
Kleihues P, Louis DN, Scheithauer BW et al (2002) The WHO classification of tumors of the nervous system. J Neuropathol Exp Neurol 61:215–225 discussion 26–29
Reya T, Morrison SJ, Clarke MF et al (2001) Stem cells, cancer, and cancer stem cells. Nature 414:105–111
Bleau AM, Hambardzumyan D, Ozawa T et al (2009) PTEN/PI3K/Akt pathway regulates the side population phenotype and ABCG2 activity in glioma tumor stem-like cells. Cell Stem Cell 4:226–235
Qiu B, Zhang D, Wang C et al (2011) IL-10 and TGF-beta2 are overexpressed in tumor spheres cultured from human gliomas. Mol Biol Rep 38(5):3585–3591
Bao S, Wu Q, McLendon RE et al (2006) Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 444:756–760
Bar EE, Chaudhry A, Lin A et al (2007) Cyclopamine-mediated hedgehog pathway inhibition depletes stem-like cancer cells in glioblastoma. Stem Cells 25:2524–2533
Zhou BB, Zhang H, Damelin M et al (2009) Tumour-initiating cells: challenges and opportunities for anticancer drug discovery. Nat Rev Drug Discov 8:806–823
Patrawala L, Calhoun T, Schneider-Broussard R et al (2005) Side population is enriched in tumorigenic, stem-like cancer cells, whereas ABCG2+ and ABCG2− cancer cells are similarly tumorigenic. Cancer Res 65:6207–6219
Politi K, Feirt N, Kitajewski J (2004) Notch in mammary gland development and breast cancer. Semin Cancer Biol 14:341–347
Weijzen S, Rizzo P, Braid M et al (2002) Activation of Notch-1 signaling maintains the neoplastic phenotype in human Ras-transformed cells. Nat Med 8:979–986
Dievart A, Beaulieu N, Jolicoeur P (1999) Involvement of Notch1 in the development of mouse mammary tumors. Oncogene 18:5973–5981
Purow BW, Haque RM, Noel MW et al (2005) Expression of notch-1 and its ligands, delta-like-1 and jagged-1, is critical for glioma cell survival and proliferation. Cancer Res 65:2353–2363
Jin F, Zhao L, Zhao HY et al (2008) Paradoxical expression of anti-apoptotic and MRP genes on cancer stem-like cell isolated from TJ905 glioblastoma multiforme cell line. Cancer Invest 26:338–343
Stupp R, Mason WP, van den Bent MJ et al (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996
Wang C, Cao S, Tie X et al (2011) Induction of cytotoxicity by photoexcitation of TiO(2) can prolong survival in glioma-bearing mice. Mol Biol Rep 38(1):523–530
Nakada M, Nakada S, Demuth T et al (2007) Molecular targets of glioma invasion. Cell Mol Life Sci 64:458–478
Singh SK, Hawkins C, Clarke ID et al (2004) Identification of human brain tumour initiating cells. Nature 432:396–401
Artavanis-Tsakonas S, Rand MD, Lake RJ (1999) Notch signaling: cell fate control and signal integration in development. Science 284:770–776
Nickoloff BJ, Osborne BA, Miele L (2003) Notch signaling as a therapeutic target in cancer: a new approach to the development of cell fate modifying agents. Oncogene 22:6598–6608
Lai EC (2002) Keeping a good pathway down: transcriptional repression of Notch pathway target genes by CSL proteins. EMBO Rep 3:840–845
Pece S, Serresi M, Santolini E et al (2004) Loss of negative regulation by numb over Notch is relevant to human breast carcinogenesis. J Cell Biol 167:215–221
Fan X, Khaki L, Zhu TS et al (2010) NOTCH pathway blockade depletes CD133-positive glioblastoma cells and inhibits growth of tumor neurospheres and xenografts. Stem Cells 28:5–16
Kanamori M, Kawaguchi T, Nigro JM et al (2007) Contribution of Notch signaling activation to human glioblastoma multiforme. J Neurosurg 106:417–427
Searfoss GH, Jordan WH, Calligaro DO et al (2003) Adipsin, a biomarker of gastrointestinal toxicity mediated by a functional gamma-secretase inhibitor. J Biol Chem 278:46107–46116
Merkerova M, Klamova H, Brdicka R et al (2007) Targeting of gene expression by siRNA in CML primary cells. Mol Biol Rep 34(1):27–33
Grzelinski M, Urban-Klein B, Martens T et al (2006) RNA interference-mediated gene silencing of pleiotrophin through polyethylenimine-complexed small interfering RNAs in vivo exerts antitumoral effects in glioblastoma xenografts. Hum Gene Ther 17:751–766
Biswal BK, Debata NB, Verma RS (2010) Development of a targeted siRNA delivery system using FOL–PEG–PEI conjugate. Mol Biol Rep 37:2919–2926
Phillips TM, McBride WH, Pajonk F (2006) The response of CD24(−/low)/CD44+ breast cancer-initiating cells to radiation. J Natl Cancer Inst 98:1777–1785
Acknowledgment
This study was supported by the Nature Science Foundation of Shandong Province, China (Y2008C58).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Wang, J., Wang, C., Meng, Q. et al. siRNA targeting Notch-1 decreases glioma stem cell proliferation and tumor growth. Mol Biol Rep 39, 2497–2503 (2012). https://doi.org/10.1007/s11033-011-1001-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-011-1001-1