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BMC Neurology
Published in: BMC Neurology 1/2016

Open Access 01-12-2016 | Research article

Chloroquine, an autophagy inhibitor, potentiates the radiosensitivity of glioma initiating cells by inhibiting autophagy and activating apoptosis

Authors: Hongxing Ye, Mantao Chen, Fei Cao, Hongguang Huang, Renya Zhan, Xiujue Zheng

Published in: BMC Neurology | Issue 1/2016

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Abstract

Background

Glioblastoma is refractory to conventional treatment, which is combined of surgery, chemotherapy and radiotherapy. Recent studies have shown that glioma initiating cells (GICs) contribute to tumorigenesis and radioresistance. Recently, other studies showed that the GICs use the autophagy as the major pathway to survive. Chloroquine, an anti-malarial chemical, is an autophagic inhibitor which blocks autophagosome fusion with lysosome and slows down lysosomal acidification. The aim of this study was to explore the mechanisms of chloroquine on the radiosensitivity of GICs.

Methods

Human glioblastoma cell lines U87 were investigated. MTT and clonogenic survival assay were used to evaluate the cell viability and survival from radiation. The formation of autophagosomes were evaluated by immunofluorescence. Annexin V-FITC/PI staining and flow cytometry were used to quantify the apoptotic cells. The expression levels of proteins were analyzed by Western blot. Cell cycle status was analyzed by checking DNA content after staining with PI. A comet assay was used to assess the DNA repair in the cells. Tumorsphere assay was used for evaluating GICs’ renewal ability.

Results

Treatment of U87 GICs with chloroquine (10–80 nmol/L) alone inhibited the cell growth in a dose-dependent manner. A dose of chloroquine (20 nmol/L) obviously enhanced the radiation sensitivity of U87 GICs., we found more punctate patterns of microtubule-associated protein LC3 immunoreactivity in radiation-treated U87 GICs, and the level of membrane-bound LC3-II was obviously enhanced. A combination of radiation and chloroquine obviously enhanced the U87 GICs’ apoptosis, as demonstrated by the enhanced levels of caspase-3, and reduced level of Bcl-2. In additon, combination of radiation and chloroquine cause G1/G0 cell cycle arrest. what’s more, Chloroquine obviously weakened the repair of radiation-induced DNA damage as reflected by the tail length of the comet. Combination treatment of irradiation and chloroquine has synergistic effects on decreasing the GICs’ tumorsphere number and diameter.

Conclusion

Chloroquine enhances the radiosensitivity of GICs in vitro, suggesting the feasibility of joint treatment with chloroquine with radiation for GBM.
Literature
1.
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987–96.CrossRefPubMed
2.
Kim SH, Ezhilarasan R, Phillips E, Gallego-Perez D, Sparks A, Taylor D, Ladner K, Furuta T, Sabit H, Chhipa R, et al. Serine/threonine kinase MLK4 determines mesenchymal identity in glioma stem cells in an NF-kappa B-dependent manner. Cancer Cell. 2016;29(2):201–13.CrossRefPubMedPubMedCentral
3.
Valent P, Bonnet D, Wohrer S, Andreeff M, Copland M, Chomienne C, Eaves C. Heterogeneity of neoplastic stem cells: theoretical, functional, and clinical implications. Cancer Res. 2013.
4.
Brown CE, Starr R, Aguilar B, Shami AF, Martinez C, D’Apuzzo M, Barish ME, Forman SJ, Jensen MC. Stem-like tumor-initiating cells isolated from IL13Ralpha2 expressing gliomas are targeted and killed by IL13-zetakine-redirected T cells. Clin Cancer Res. 2012;18(8):2199–209.CrossRefPubMedPubMedCentral
5.
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, Dewhirst MW, Bigner DD, Rich JN. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006;444(7120):756–60.CrossRefPubMed
6.
Choi AM, Ryter SW, Levine B. Autophagy in human health and disease. N Engl J Med. 2013;368(19):1845–6.CrossRefPubMed
7.
Zois CE, Koukourakis MI. Radiation-induced autophagy in normal and cancer cells: towards novel cytoprotection and radio-sensitization policies? Autophagy. 2009;5(4):442–50.CrossRefPubMed
8.
Yao KC, Komata T, Kondo Y, Kanzawa T, Kondo S, Germano IM. Molecular response of human glioblastoma multiforme cells to ionizing radiation: cell cycle arrest, modulation of the expression of cyclin-dependent kinase inhibitors, and autophagy. J Neurosurg. 2003;98(2):378–84.CrossRefPubMed
9.
Koukourakis MI, Mitrakas AG, Giatromanolaki A. Therapeutic interactions of autophagy with radiation and temozolomide in glioblastoma: evidence and issues to resolve. Br J Cancer. 2016;114(5):485–96.CrossRefPubMedPubMedCentral
10.
Pascolo S. Time to use a dose of Chloroquine as an adjuvant to anti-cancer chemotherapies. Eur J Pharmacol. 2016;771:139–44.CrossRefPubMed
11.
Chen TY, Syu JS, Lin TC, Cheng HL, Lu FL, Wang CY. Chloroquine alleviates etoposide-induced centrosome amplification by inhibiting CDK2 in adrenocortical tumor cells. Oncogenesis. 2015;4:e180.CrossRefPubMedPubMedCentral
12.
Rosenfeld MR, Ye X, Supko JG, Desideri S, Grossman SA, Brem S, Mikkelson T, Wang D, Chang YC, Hu J, et al. A phase I/II trial of hydroxychloroquine in conjunction with radiation therapy and concurrent and adjuvant temozolomide in patients with newly diagnosed glioblastoma multiforme. Autophagy. 2014;10(8):1359–68.CrossRefPubMedPubMedCentral
13.
Briceno E, Calderon A, Sotelo J. Institutional experience with chloroquine as an adjuvant to the therapy for glioblastoma multiforme. Surg Neurol. 2007;67(4):388–91.CrossRefPubMed
14.
Bulut T, Ekici MA, Tucer B, Basarslan SK, Kamasak K, Kurtsoy A. The effect of chloroquine treatment in malignant astrocytomas on prognosis. Turk J Med Sci. 2013;43(5):782–9.CrossRef
15.
Wang WJ, Long LM, Yang N, Zhang QQ, Ji WJ, Zhao JH, Qin ZH, Wang Z, Chen G, Liang ZQ. NVP-BEZ235, a novel dual PI3K/mTOR inhibitor, enhances the radiosensitivity of human glioma stem cells in vitro. Acta Pharmacol Sin. 2013;34(5):681–90.CrossRefPubMedPubMedCentral
16.
Gong C, Bauvy C, Tonelli G, Yue W, Delomenie C, Nicolas V, Zhu Y, Domergue V, Marin-Esteban V, Tharinger H, et al. Beclin 1 and autophagy are required for the tumorigenicity of breast cancer stem-like/progenitor cells. Oncogene. 2013;32(18):2261–72. 2272e 2261–2211.CrossRefPubMed
17.
Berardi DE, Flumian C, Rodriguez CE, Bessone MID, Cirigliano SM, Joffe EDBD, Fiszman GL, Urtreger AJ, Todaro LB. PKC delta inhibition impairs mammary cancer proliferative capacity but selects cancer stem cells, involving autophagy. J Cell Biochem. 2016;117(3):730–40.CrossRefPubMed
18.
Yu VY, Nguyen D, Pajonk F, Kupelian P, Kaprealian T, Selch M, Low DA, Sheng K. Incorporating cancer stem cells in radiation therapy treatment response modeling and the implication in glioblastoma multiforme treatment resistance. Int J Radiat Oncol Biol Phys. 2015;91(4):866–75.CrossRefPubMed
19.
Shen Y, Chen H, Zhang J, Chen Y, Wang M, Ma J, Hong L, Liu N, Fan Q, Lu X, et al. Increased notch signaling enhances radioresistance of malignant stromal cells induced by glioma stem/progenitor cells. PLoS One. 2015;10(11):e0142594.CrossRefPubMedPubMedCentral
20.
21.
Cory S, Adams JM. The BCL2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer. 2002;2(9):647–56.CrossRefPubMed
22.
Sotelo J, Briceno E, Lopez-Gonzalez MA. Adding chloroquine to conventional treatment for glioblastoma multiforme: a randomized, double-blind, placebo-controlled trial. Ann Intern Med. 2006;144(5):337–43.CrossRefPubMed
23.
Bilger A, Bittner MI, Grosu AL, Wiedenmann N, Meyer PT, Firat E, Niedermann G, Weber WA, Milanovic D. FET-PET-based reirradiation and chloroquine in patients with recurrent glioblastoma. Strahlenther Onkol. 2014;190(10):957–61.CrossRefPubMed
24.
Chen P, Hu T, Liang YP, Jiang YN, Pan YF, Li CJ, Zhang P, Wei DP, Li P, Jeong LS, et al. Synergistic inhibition of autophagy and neddylation pathways as a novel therapeutic approach for targeting liver cancer. Oncotarget. 2015;6(11):9002–17.CrossRefPubMedPubMedCentral
25.
Sehgal AR, Konig H, Johnson DE, Tang D, Amaravadi RK, Boyiadzis M, Lotze MT. You eat what you are: autophagy inhibition as a therapeutic strategy in leukemia. Leukemia. 2015;29(3):517–25.CrossRefPubMed
26.
Yuan XP, Du J, Hua S, Zhang HW, Gu C, Wang J, Yang L, Huang JF, Yu JH, Liu FJ. Suppression of autophagy augments the radiosensitizing effects of STAT3 inhibition on human glioma cells. Exp Cell Res. 2015;330(2):267–76.CrossRefPubMed
27.
Ye F, Zhang YB, Liu Y, Yamada K, Tso JL, Menjivar JC, Tian JY, Yong WH, Schaue D, Mischel PS, et al. Protective properties of radio-chemoresistant glioblastoma stem cell clones are associated with metabolic adaptation to reduced glucose dependence. PLoS One. 2013;8(11):e80397.CrossRefPubMedPubMedCentral
28.
Kim H, Bernard M, Flickinger J, Epperly MW, Wang H, Dixon TM, Shields D, Houghton F, Greenberger JS. The autophagy inducing drug carbamazepine is a radiation protector and mitigator. Cancer Res. 2011;71.
29.
Firat E, Weyerbrock A, Gaedicke S, Grosu AL, Niedermann G. Chloroquine or chloroquine-PI3K/Akt pathway inhibitor combinations strongly promote gamma-irradiation-induced cell death in primary stem-like glioma cells. PLoS One. 2012;7(10):e47357.CrossRefPubMedPubMedCentral
Metadata
Title
Chloroquine, an autophagy inhibitor, potentiates the radiosensitivity of glioma initiating cells by inhibiting autophagy and activating apoptosis
Authors
Hongxing Ye
Mantao Chen
Fei Cao
Hongguang Huang
Renya Zhan
Xiujue Zheng
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Neurology / Issue 1/2016
Electronic ISSN: 1471-2377
DOI
https://doi.org/10.1186/s12883-016-0700-6

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