Cellular and Molecular Neurobiology

, Volume 36, Issue 7, pp 1197–1208 | Cite as

Inhibition of Autophagy by Chloroquine Enhances the Antitumor Efficacy of Sorafenib in Glioblastoma

  • Xiangyu Liu
  • Kangjian Sun
  • Handong Wang
  • Yuyuan DaiEmail author
Original Research


Glioblastoma multiforme (GBM) is the most aggressive and common brain tumor in adults. Sorafenib, a multi-kinase inhibitor, has been shown to inhibit cell proliferation and induce apoptosis through inhibition of STAT3 signaling in glioblastoma cells and in intracranial gliomas. However, sorafenib also induces cell autophagy. Due to the dual roles of autophagy in tumor cell survival and death, the therapeutic effect of sorafenib on glioblastoma is uncertain. Here, we combined sorafenib treatment in GBM cells (U373 and LN229) and tumors with the autophagy inhibitor chloroquine. We found that blockage of autophagy further inhibited cell proliferation and migration and induced cell apoptosis in vitro and in vivo. These findings suggest the possibility of combination treatment with sorafenib and autophagy inhibitors for GBM.


Glioblastoma Sorafenib Autophagy Apoptosis 



This work was supported by Grants from Nanjing Jinling Hospital, Nanjing, China (2011Q047).

Supplementary material

10571_2015_318_MOESM1_ESM.tiff (436 kb)
Fig. S1 Sorafenib and chloroquine cooperated to inhibit the glioma cell viability. Human glioma cell lines LN229 (black line) and U373 (grey line) were plated in 96-well plates (5×103/well), and treated with sorafenib alone (10 µM) and in combination with 5 µM chloroquine for 24, 48 and 72 hours. Cell viability was examined by CCK-8 assay. Relative viability showed that sorafenib and chloroquine cooperate to promote cell death. Each bar represents mean ± SD of triplicate determinations; results shown are representative of three identical experiments.* p <0.05 vs. cell viability at 0 µM sorafenib in each groups (TIFF 438 kb)
10571_2015_318_MOESM2_ESM.tif (10 mb)
Fig. S2 Inhibition of autophagy with ATG5 siRNA enhanced the inhibitory activity of sorafenib on gliomas cell viability. A. Human glioma cell line LN229 and U373 were plated in 6-well plates (1×106/well), and treated with control siRNA, siATG5 sequence 1 and sequence 2 for 48 hours. 10 µg of cell lysate was analyzed by Western blot using ATG5 and β-actin antibodies. B. Human glioma cell lines LN229 and U373 were plated in 6-well plates (1×106/well), and treated with control siRNA (siControl) and siRNA (siATG5) sequence 2 for 12 hours. Then the cells were trypsin digested and were plated in 96-well plates (5×103/well), and treated with 0-20 µM sorafenib for 48 hours. Cell viability was examined by CCK-8 assay. Relative viability showed that sorafenib and chloroquine cooperate to promote cell death. Each bar represents mean ± SD of triplicate determinations; results shown are representative of three identical experiments.* p <0.05 vs. cell viability at 0 µM sorafenib in each groups (TIFF 10,231 kb)
10571_2015_318_MOESM3_ESM.tif (9 mb)
Fig. S3 Cell viability of LN229 and U373 treated with sorafenib alone (2.5 µM) and in combination with 5 µM chloroquine for 24 hours. 0.1% DMSO was used as control. Each bar represents mean ± SD of triplicate determinations; results shown are representative of three identical experiments. No statistical difference were found between different groups (TIFF 9,186 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Xiangyu Liu
    • 1
  • Kangjian Sun
    • 1
  • Handong Wang
    • 1
  • Yuyuan Dai
    • 2
    Email author
  1. 1.Department of Neurosurgery, Nanjing Jinling Hospital, School of MedicineNanjing UniversityNanjingPeople’s Republic of China
  2. 2.Department of Surgery, School of MedicineUniversity of CaliforniaSan FranciscoUSA

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