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Targeting autophagy enhances BO-1051-induced apoptosis in human malignant glioma cells

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Abstract

Purpose

BO-1051 is an N-mustard derivative that is conjugated with DNA-affinic 9-anilinoacridine. Since BO-1051 was reported to have strong anticancer activity, we investigated the effect and underlying mechanism of BO-1051 in human glioma cell lines.

Methods

Human glioma cell lines U251MG and U87MG were studied with BO-1051 or the combination of BO-1051 and autophagic inhibitors. Growth inhibition was assessed by MTT assay. Apoptosis was measured by annexin V staining followed by flow cytometry and immunoblotting for apoptosis-related molecules. Induction of autophagy was detected by acridine orange labeling, electron microscopy, LC3 localization and its conversion. Transfection of shRNA was used to determine the involvement of Beclin1 in apoptotic cell death.

Results

MTT assay showed that BO-1051 suppressed the viability of four glioma cell lines (U251MG, U87MG, GBM-3 and DBTRG-05MG) in a dose-dependent manner. The IC50 values of BO-1051 for the glioma cells were significantly lower than the values for primary neurons cultures and normal fibroblast cells. Moreover, BO-1051 not only induced apoptotic cell death, but also enhanced autophagic flux via inhibition of Akt/mTOR and activation of Erk1/2. Importantly, suppression of autophagy by 3-methyladenine or bafilomycin A1 significantly increased BO-1051-induced apoptotic cell death in U251MG and U87MG cells. In addition, the proportion of apoptotic cells after BO-1051 treatment was enhanced by co-treatment with shRNA against Beclin1.

Conclusions

BO-1051 induced both apoptosis and autophagy, and inhibition of autophagy significantly augmented the cytotoxic effect of BO-1051. Thus, a combination of BO-1051 and autophagic inhibitors offers a potentially new therapeutic modality for the treatment of malignant glioma.

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Acknowledgments

This study was supported by research grants from the National Science Council (NSC97-3111-B-075-001-MY3, NSC98-2320-B-075-003-MY3, NSC99-2628-B- 016-014-MY3 and NSC99-2811-B-016-007-MY3), Taipei Veterans General Hospital (V97B1-006 and E1-008, F-001), Tri-Service General Hospital (TSGH-C100-047), the Joint Projects of UTVGH (VGHUST 98-p1-01), Yen-Tjing-Ling Medical Foundation (96/97/98), National Yang-Ming University (Ministry of Education, Aim for the Top University Plan) & Genomic Center Project, Institute of Biological medicine, Academia Sinica (IBMS-CRC99-p01), and Center of Excellence for Cancer Research at Taipei Veterans General Hospital (DOH99-TD-C-111-007), Taiwan.

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Graduate Institutes of Life Sciences, National Defense Medical Center, Taipei, Taiwan, ROC

    Pei-Ming Chu & Hsin-I Ma

  2. Department of Neurological Surgery, Tri-Service General Hospital, No. 325, Sec. 2, Cheng-Kung Road, Taipei, 114, Taiwan, ROC

    Pei-Ming Chu & Hsin-I Ma

  3. Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan

    Li-Hsin Chen & Shih-Hwa Chiou

  4. Department of Neurosurgery, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan

    Ming-Teh Chen & Yang-Hsin Shih

  5. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    Tsann-Long Su & Pang-Hsien Tu

  6. Department of Medical Research and Education, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei, 112, Taiwan, ROC

    Pei-Chen Hsieh, Yu-Chih Chen & Shih-Hwa Chiou

  7. Institute of Oral Biology, National Yang-Ming University, Taipei, Taiwan

    Chian-Shiu Chien & Bo-Hua Jiang

  8. School of Pharmacy, China Medical University, Taichung, Taiwan

    Yi-Hui Lin

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  1. Pei-Ming Chu
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Correspondence to Hsin-I Ma or Shih-Hwa Chiou.

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Chu, PM., Chen, LH., Chen, MT. et al. Targeting autophagy enhances BO-1051-induced apoptosis in human malignant glioma cells. Cancer Chemother Pharmacol 69, 621–633 (2012). https://doi.org/10.1007/s00280-011-1747-0

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  • DOI: https://doi.org/10.1007/s00280-011-1747-0

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