Chemotherapy is an adjuvant treatment for glioblastomas, however, chemotherapy remains palliative because of the development of multidrug resistance (MDR). Following prolonged chemotherapy, MDR protein 1 (MDR1) and CD133 increase in recurrent glioblastomas. CD133 positive (CD133+) glioma cancer stem-like cells (GCSCs) markedly promote drug resistance and exhibit increased DNA damage repair capability; thus they have a key role in determining tumor chemosensitivity. Although CD133, DNA-dependent protein kinase (DNA-PK), and MDR1 are elevated in CD133+ GCSCs, the relationship among these molecules has not been elucidated. In this study, MDR glioblastoma cell lines were created in response to prolonged doxorubicin chemotherapy. CD133, DNA-PK and MDR1 were markedly elevated in these cells. CD133 and DNA-PK may increase MDR1 via the phosphatidylinositol-3-kinase (PI3K)-Akt signal pathway. PI3K downstream targets Akt and nuclear factor (NF)-κB, which interacts with the MDR1 promoter, were also elevated in these cells. Downregulation of CD133 and DNA-PK by small interfering RNA, or inhibition of PI3K or Akt, decreased Akt, NF-κB and MDR1 expression. The results indicate that CD133 and DNA-PK regulate MDR1 through the PI3K- or Akt-NF-κB signal pathway. Consequently, a novel chemotherapeutic regimen targeting CD133 and DNA-PK in combination with traditional protocols may increase chemotherapeutic efficacy and improve prognosis for individuals who present with glioblastoma.
Subscribe to Journal
Get full journal access for 1 year
only $51.94 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC et al. Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 2009; 10: 459–466.
Stupp R, Hegi ME, van den Bent MJ, Mason WP, Weller M, Mirimanoff RO et al. Changing paradigms—an update on the multidisciplinary management of malignant glioma. Oncologist 2006; 11: 165–180.
Hegi ME, Diserens AC, Gorlia T, Hamou MF, de Tribolet N, Weller M et al. MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med 2005; 352: 997–1003.
Gottesman MM, Fojo T, Bates SE . Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2002; 2: 48–58.
Abe T, Hasegawa S, Taniguchi K, Yokomizo A, Kuwano T, Ono M et al. Possible involvement of multidrug-resistance-associated protein (MRP) gene expression in spontaneous drug resistance to vincristine, etoposide and adriamycin in human glioma cells. Int J Cancer 1994; 58: 860–864.
Feun LG, Savaraj N, Landy HJ . Drug resistance in brain tumors. J Neurooncol 1994; 20: 165–176.
de Faria GP, de Oliveira JA, de Oliveira JG, Romano Sde O, Neto VM, Maia RC . Differences in the expression pattern of P-glycoprotein and MRP1 in low-grade and high-grade gliomas. Cancer Invest 2008; 26: 883–889.
Borst P, Schinkel AH . P-glycoprotein ABCB1: a major player in drug handling by mammals. J Clin Invest 2013; 123: 4131–4133.
Matsumoto T, Tani E, Kaba K, Shindo H, Miyaji K . Expression of P-glycoprotein in human glioma cell lines and surgical glioma specimens. J Neurosurg 1991; 74: 460–466.
Bahr O, Rieger J, Duffner F, Meyermann R, Weller M, Wick W . P-glycoprotein and multidrug resistance-associated protein mediate specific patterns of multidrug resistance in malignant glioma cell lines, but not in primary glioma cells. Brain Pathol 2003; 13: 482–494.
Decleves X, Fajac A, Lehmann-Che J, Tardy M, Mercier C, Hurbain I et al. Molecular and functional MDR1-Pgp and MRPs expression in human glioblastoma multiforme cell lines. Int J Cancer 2002; 98: 173–180.
Robey RW, Polgar O, Deeken J, To KW, Bates SE . ABCG2: determining its relevance in clinical drug resistance. Cancer Metastasis Rev 2007; 26: 39–57.
Jordan CT, Guzman ML, Noble M . Cancer stem cells. N Engl J Med 2006; 355: 1253–1261.
Tamura K, Aoyagi M, Ando N, Ogishima T, Wakimoto H, Yamamoto M et al. Expansion of CD133-positive glioma cells in recurrent de novo glioblastomas after radiotherapy and chemotherapy. J Neurosurg 2013; 119: 1145–1155.
Liu Q, Nguyen DH, Dong Q, Shitaku P, Chung K, Liu OY et al. Molecular properties of CD133+ glioblastoma stem cells derived from treatment-refractory recurrent brain tumors. J Neurooncol 2009; 94: 1–19.
Liu G, Yuan X, Zeng Z, Tunici P, Ng H, Abdulkadir IR et al. Analysis of gene expression and chemoresistance of CD133+ cancer stem cells in glioblastoma. Mol Cancer 2006; 5: 67.
Angelastro JM, Lame MW . Overexpression of CD133 promotes drug resistance in C6 glioma cells. Mol Cancer Res 2010; 8: 1105–1115.
Hennessy BT, Smith DL, Ram PT, Lu Y, Mills GB . Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat Rev Drug Disc 2005; 4: 988–1004.
West KA, Castillo SS, Dennis PA . Activation of the PI3K/Akt pathway and chemotherapeutic resistance. Drug Resist Updates 2002; 5: 234–248.
Hambardzumyan D, Squatrito M, Carbajal E, Holland EC . Glioma formation, cancer stem cells, and akt signaling. Stem Cell Rev 2008; 4: 203–210.
Kao GD, Jiang Z, Fernandes AM, Gupta AK, Maity A . Inhibition of phosphatidylinositol-3-OH kinase/Akt signaling impairs DNA repair in glioblastoma cells following ionizing radiation. J Biol Chem 2007; 282: 21206–21212.
Choi BH, Kim CG, Lim Y, Shin SY, Lee YH . Curcumin down-regulates the multidrug-resistance mdr1b gene by inhibiting the PI3K/Akt/NF kappa B pathway. Cancer Lett 2008; 259: 111–118.
Lin X, Zhang X, Wang Q, Li J, Zhang P, Zhao M et al. Perifosine downregulates MDR1 gene expression and reverses multidrug-resistant phenotype by inhibiting PI3K/Akt/NF-kappaB signaling pathway in a human breast cancer cell line. Neoplasma 2012; 59: 248–256.
Barancik M, Bohacova V, Sedlak J, Sulova Z, Breier A . LY294,002, a specific inhibitor of PI3K/Akt kinase pathway, antagonizes P-glycoprotein-mediated multidrug resistance. Eur J Pharm Sci 2006; 29: 426–434.
Tazzari PL, Cappellini A, Ricci F, Evangelisti C, Papa V, Grafone T et al. Multidrug resistance-associated protein 1 expression is under the control of the phosphoinositide 3 kinase/Akt signal transduction network in human acute myelogenous leukemia blasts. Leukemia 2007; 21: 427–438.
Kuo MT, Liu Z, Wei Y, Lin-Lee YC, Tatebe S, Mills GB et al. Induction of human MDR1 gene expression by 2-acetylaminofluorene is mediated by effectors of the phosphoinositide 3-kinase pathway that activate NF-kappaB signaling. Oncogene 2002; 21: 1945–1954.
Wei Y, Jiang Y, Zou F, Liu Y, Wang S, Xu N et al. Activation of PI3K/Akt pathway by CD133-p85 interaction promotes tumorigenic capacity of glioma stem cells. Proc Natl Acad Sci USA 2013; 110: 6829–6834.
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 2006; 444: 756–760.
Facchino S, Abdouh M, Chatoo W, Bernier G . BMI1 confers radioresistance to normal and cancerous neural stem cells through recruitment of the DNA damage response machinery. J Neurosci 2010; 30: 10096–10111.
Lee PC, Lee HJ, Kakadiya R, Sanjiv K, Su TL, Lee TC . Multidrug-resistant cells overexpressing P-glycoprotein are susceptible to DNA crosslinking agents due to attenuated Src/nuclear EGFR cascade-activated DNA repair activity. Oncogene 2013; 32: 1144–1154.
Bolderson E, Richard DJ, Zhou BB, Khanna KK . Recent advances in cancer therapy targeting proteins involved in DNA double-strand break repair. Clin Cancer Res 2009; 15: 6314–6320.
Mould E, Berry P, Jamieson D, Hill C, Cano C, Tan N et al. Identification of dual DNA-PK MDR1 inhibitors for the potentiation of cytotoxic drug activity. Biochem Pharmacol 2014; 88: 58–65.
Lees-Miller SP, Godbout R, Chan DW, Weinfeld M, Day RS 3rd, Barron GM et al. Absence of p350 subunit of DNA-activated protein kinase from a radiosensitive human cell line. Science 1995; 267: 1183–1185.
Seo SB, Hur JG, Kim MJ, Lee JW, Kim HB, Bae JH et al. TRAIL sensitize MDR cells to MDR-related drugs by down-regulation of P-glycoprotein through inhibition of DNA-PKcs/Akt/GSK-3beta pathway and activation of caspases. Mol Cancer 2010; 9: 199.
Shimura T, Noma N, Oikawa T, Ochiai Y, Kakuda S, Kuwahara Y et al. Activation of the AKT/cyclin D1/Cdk4 survival signaling pathway in radioresistant cancer stem cells. Oncogenesis 2012; 1: e12.
Basu S, Rosenzweig KR, Youmell M, Price BD . The DNA-dependent protein kinase participates in the activation of NF kappa B following DNA damage. Biochem Biophys Res Commun 1998; 247: 79–83.
Baeuerle PA, Baltimore D . I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science 1988; 242: 540–546.
Marquardt D, Center MS . Drug transport mechanisms in HL60 cells isolated for resistance to adriamycin: evidence for nuclear drug accumulation and redistribution in resistant cells. Cancer Res 1992; 52: 3157–3163.
Keizer HG, Schuurhuis GJ, Broxterman HJ, Lankelma J, Schoonen WG, van Rijn J et al. Correlation of multidrug resistance with decreased drug accumulation, altered subcellular drug distribution, and increased P-glycoprotein expression in cultured SW-1573 human lung tumor cells. Cancer Res 1989; 49: 2988–2993.
Barr MP, Gray SG, Hoffmann AC, Hilger RA, Thomale J, O'Flaherty JD et al. Generation and characterisation of Cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. PloS One 2013; 8: e54193.
Liu YP, Yang CJ, Huang MS, Yeh CT, Wu AT, Lee YC et al. Cisplatin selects for multidrug-resistant CD133+ cells in lung adenocarcinoma by activating notch signaling. Cancer Res 2013; 73: 406–416.
Freitas DP, Teixeira CA, Santos-Silva F, Vasconcelos MH, Almeida GM . Therapy-induced enrichment of putative lung cancer stem-like cells. Int J Cancer 2014; 134: 1270–1278.
Jin X, Jin X, Jung JE, Beck S, Kim H . Cell surface Nestin is a biomarker for glioma stem cells. Biochem Biophys Res Commun 2013; 433: 496–501.
Guo Y, Liu S, Wang P, Zhao S, Wang F, Bing L et al. Expression profile of embryonic stem cell-associated genes Oct4, Sox2 and Nanog in human gliomas. Histopathology 2011; 59: 763–775.
Ma L, Lai D, Liu T, Cheng W, Guo L . Cancer stem-like cells can be isolated with drug selection in human ovarian cancer cell line SKOV3. Acta Biochim Biophys Sin (Shanghai) 2010; 42: 593–602.
Bertolini G, Roz L, Perego P, Tortoreto M, Fontanella E, Gatti L et al. Highly tumorigenic lung cancer CD133+ cells display stem-like features and are spared by cisplatin treatment. Proc Natl Acad Sci USA 2009; 106: 16281–16286.
Liu T, Xu F, Du X, Lai D, Liu T, Zhao Y et al. Establishment and characterization of multi-drug resistant, prostate carcinoma-initiating stem-like cells from human prostate cancer cell lines 22RV1. Mol Cell Biochem 2010; 340: 265–273.
Frosina G . DNA repair and resistance of gliomas to chemotherapy and radiotherapy. Mol Cancer Res 2009; 7: 989–999.
Hsu FM, Zhang S, Chen BP . Role of DNA-dependent protein kinase catalytic subunit in cancer development and treatment. Transl Cancer Res 2012; 1: 22–34.
Abe T, Mori T, Wakabayashi Y, Nakagawa M, Cole SP, Koike K et al. Expression of multidrug resistance protein gene in patients with glioma after chemotherapy. J Neurooncol 1998; 40: 11–18.
Mirkin BL, Clark S, Zheng X, Chu F, White BD, Greene M et al. Identification of midkine as a mediator for intercellular transfer of drug resistance. Oncogene 2005; 24: 4965–4974.
Mannervik B, Guthenberg C . Glutathione transferase (human placenta). Methods Enzymol 1981; 77: 231–235.
Fairchild CR, Ivy SP, Kao-Shan CS, Whang-Peng J, Rosen N, Israel MA et al. Isolation of amplified and overexpressed DNA sequences from adriamycin-resistant human breast cancer cells. Cancer Res 1987; 47: 5141–5148.
Van der Bliek AM, Baas F, Van der Velde-Koerts T, Biedler JL, Meyers MB, Ozols RF et al. Genes amplified and overexpressed in human multidrug-resistant cell lines. Cancer Res 1988; 48: 5927–5932.
Nakai E, Park K, Yawata T, Chihara T, Kumazawa A, Nakabayashi H et al. Enhanced MDR1 expression and chemoresistance of cancer stem cells derived from glioblastoma. Cancer Invest 2009; 27: 901–908.
Huet S, Schott B, Robert J . P-glycoprotein overexpression cannot explain the complete doxorubicin-resistance phenotype in rat glioblastoma cell lines. Br J Cancer 1992; 65: 538–544.
Bleau AM, Huse JT, Holland EC . The ABCG2 resistance network of glioblastoma. Cell Cycle (Georgetown, TX) 2009; 8: 2936–2944.
Dean M, Fojo T, Bates S . Tumour stem cells and drug resistance. Nat Rev Cancer 2005; 5: 275–284.
Balik V, Mirossay P, Bohus P, Sulla I, Mirossay L, Sarissky M . Flow cytometry analysis of neural differentiation markers expression in human glioblastomas may predict their response to chemotherapy. Cell Mol Neurobiol 2009; 29: 845–858.
Zhong X, Safa AR . Phosphorylation of RNA helicase A by DNA-dependent protein kinase is indispensable for expression of the MDR1 gene product P-glycoprotein in multidrug-resistant human leukemia cells. Biochemistry (Mosc) 2007; 46: 5766–5775.
Tacar O, Sriamornsak P, Dass CR . Doxorubicin: an update on anticancer molecular action, toxicity and novel drug delivery systems. J Pharm Pharmacol 2013; 65: 157–170.
Ramaswamy S, Nakamura N, Vazquez F, Batt DB, Perera S, Roberts TM et al. Regulation of G1 progression by the PTEN tumor suppressor protein is linked to inhibition of the phosphatidylinositol 3-kinase/Akt pathway. Proc Natl Acad Sci USA 1999; 96: 2110–2115.
Ballard DW, Dixon EP, Peffer NJ, Bogerd H, Doerre S, Stein B et al. The 65-kDa subunit of human NF-kappa B functions as a potent transcriptional activator and a target for v-Rel-mediated repression. Proc Natl Acad Sci USA 1992; 89: 1875–1879.
Skovsgaard T . Mechanisms of resistance to daunorubicin in Ehrlich ascites tumor cells. Cancer Res 1978; 38: 1785–1791.
Dano K . Active outward transport of daunomycin in resistant Ehrlich ascites tumor cells. Biochim Biophys Acta 1973; 323: 466–483.
Hussein D, Punjaruk W, Storer LC, Shaw L, Othman R, Peet A et al. Pediatric brain tumor cancer stem cells: cell cycle dynamics, DNA repair, and etoposide extrusion. Neuro-Oncology 2011; 13: 70–83.
Xi G, Robinson E, Mania-Farnell B, Vanin EF, Shim KW, Takao T et al. Convection-enhanced delivery of nanodiamond drug delivery platforms for intracranial tumor treatment. Nanomed Nanotechnol Biol Med 2014; 10: 381–391.
We are grateful to Dr Hao Luo for helpful suggestions and Drs Nimrod Miller and Yuping Derek Li for critical reading this article. This project was supported by the Rory David Deutsch Foundation, the Surgical Neuro-oncology Research Fund of Ann & Robert H Lurie Children’s Hospital (A&RLCH) of Chicago, and the Dr Ralph and Marian C Falk Medical Research Trust.
The authors declare no conflict of interest.
Supplementary Information accompanies this paper on the Oncogene website
About this article
Cite this article
Xi, G., Hayes, E., Lewis, R. et al. CD133 and DNA-PK regulate MDR1 via the PI3K- or Akt-NF-κB pathway in multidrug-resistant glioblastoma cells in vitro. Oncogene 35, 241–250 (2016) doi:10.1038/onc.2015.78
CD163, a novel therapeutic target, regulates the proliferation and stemness of glioma cells via casein kinase 2
Biochimica et Biophysica Acta (BBA) - Reviews on Cancer (2019)
Journal of Cellular and Molecular Medicine (2019)
Journal of Cellular Physiology (2019)